Cyclosporin analogues for preventing or treating hepatitis C infection

ABSTRACT

The present invention relates to cyclosporin analogues having antiviral activity against HCV and useful in the treatment of HCV infections. More particularly, the invention relates to novel cyclosporin analogue compounds, compositions containing such compounds and methods for using the same, as well as processes for making such compounds.

RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application No.61/148,583, filed on Jan. 30, 2009. The entire teachings of the aboveapplication are incorporated herein by reference.

TECHNICAL FIELD

The present invention relates to novel cyclosporin analogues havingantiviral activity against HCV and useful in the treatment of HCVinfections. More particularly, the invention relates to novelcyclosporin analogue compounds, compositions containing such compoundsand methods for using the same, as well as processes for making suchcompounds.

BACKGROUND OF THE INVENTION

Infection with HCV is a major cause of human liver disease throughoutthe world. In the US, an estimated 4.5 million Americans are chronicallyinfected with HCV. Although only 30% of acute infections aresymptomatic, greater than 85% of infected individuals develop chronic,persistent infection. Treatment costs for HCV infection have beenestimated at $5.46 billion for the US in 1997. Worldwide over 200million people are estimated to be infected chronically. HCV infectionis responsible for 40-60% of all chronic liver disease and 30% of allliver transplants. Chronic HCV infection accounts for 30% of allcirrhosis, end-stage liver disease, and liver cancer in the U.S. The CDCestimates that the number of deaths due to HCV will minimally increaseto 38,000/year by the year 2010.

There are considerable barriers to the development of anti-HCVtherapeutics, which include, but are not limited to, the persistence ofthe virus, the genetic diversity of the virus during replication in thehost, the high incident rate of the virus developing drug-resistantmutants, and the lack of reproducible infectious culture systems andsmall-animal models for HCV replication and pathogenesis. In a majorityof cases, given the mild course of the infection and the complex biologyof the liver, careful consideration must be given to antiviral drugs,which are likely to have significant side effects.

Due to the high degree of variability in the viral surface antigens,existence of multiple viral genotypes, and demonstrated specificity ofimmunity, the development of a successful vaccine in the near future isunlikely. Only two approved therapies for HCV infection are currentlyavailable. The original treatment regimen generally involves a 3-12month course of intravenous interferon-α (IFN-α), while a new approvedsecond-generation treatment involves co-treatment with IFN-α and thegeneral antiviral nucleoside mimics like ribavirin. Both of thesetreatments suffer from interferon related side effects as well as lowefficacy against HCV infections. There exists a need for the developmentof effective antiviral agents for treatment of HCV infection due to thepoor tolerability and disappointing efficacy of existing therapies.

Cyclosporin A (CsA), a neutral cyclic undecapeptide isolated from thefungus Tolypocladium injlaturn and currently marketed as Neoral andsandimmunem (Novartis, Basel, Switzerland), has been widely used for theprevention of organ transplant rejection. The molecular basis for theimmunosuppressant activity of cyclosporin A and cyclosporin analoguesbegins with the passive diffusion of the cyclosporin (Cs) molecule intothe cell, followed by binding to its intracellular receptor, cyclophilinA (CypA). CypA belongs to a family of proteins that catalyze cis-transpeptidyl-prolyl isomerization, i.e., PPIase, a rate-limiting step inprotein folding. CsA and other cyclosporin analogues bind to the activesite of CypA. However, immunosuppression is not believed to be due tothe inhibition of CypA PPIase activity. The target of the CsA-CypAcomplex is a Ca²⁺-calmodulin-dependent serine-threonine-specific proteinphosphatase, calcineurin. In T-cells responding to antigen presentation,an increase in intracellular Ca²⁺ activates calcineurin, whichsubsequently dephosphorylates the transcription factor called thenuclear factor of activated T-cells (“NFAT”). Dephosphorylated NFATundergoes a molecular change, e.g., homodimerization that allows it tocross into the nucleus, and promotes the expression of T-cell activationgenes. CsA and other immunosuppressive cyclosporin derivatives inhibitcalcineurin which results in the inhibition of expression of cytokinegenes, e.g., interleukin-2 (IL-2) that promotes T-cell activation andproliferation, i.e., immunosuppressive activity.

Cyclosporin A and certain derivatives have been reported as havinganti-HCV activity, see Watashi et al., Hepatology, 2003, Volume 38, pp1282-1288, Nakagawa et al., Biochem. Biophys. Res. Commun. 2004, Volume3 13, pp 42-7, and Shimotohno and K. Watashi, 2004 American TransplantCongress, Abstract No. 648 (American Journal of Transplantation 2004,Volume 4, Issue s8, Pages 1-653). The authors of the Nakagawa et al.paper state that certain chaperone activities, such as those ofcyclophilins, may be crucial for the processing and maturation of theviralproteins and for viral replication. Cyclosporin derivatives havingHCV activity are known from International Publication No's. WO2005/021028, WO 2006/039668, WO 2006/038088, WO 2006/039688, WO2007/112352, WO 2007/112357, WO 2007/112345, and WO 2007/041631.

A subsequent controlled clinical trial showed that a combination ofcyclosporin A with interferon α2b is more effective than interferonmonotherapy, especially in patients with high viral loads (Inoue et al.,“Combined Interferon α2b nd Cyclosporin A in the Treatment of ChronicHepatitis C: Controlled Trial,” J. Gastroenterol. 38:567-572 (2003)).

PCT International Patent Publication No. WO 2006/005610 recentlydescribed the use of a combination of cyclosporin A and pegylatedinterferon for treating hepatitis C viral infection. In addition, PCTInternational Patent Publication No. WO 2005/021028 relates to the useof non-immunosuppressive cyclosporine for treatment of HCV disorders.Also, Paeshuyse et al., “Potent and Selective Inhibition of Hepatitis CVirus Replication by the Non-Immunosuppressive Cyclosporin AnalogueDEBIO-025,” Antiviral Research 65(3):A41 (2005) recently publishedresults for a non-immunosuppressive cyclosporin analogue, DEBIO-025,that exhibited potent and selective inhibition of hepatitis C virusreplication. Debio-025 does possess potent binding affinity forcyclophilin A.

SUMMARY OF THE INVENTION

The present invention relates to cyclosporin analogues representedherein below, pharmaceutical compositions comprising such compounds, andmethods for the treatment of viral (particularly hepstitis C viral)infection in a subject in need of such therapy with said compounds.

In its principal embodiment, the present invention provides a compoundof formula (I);

where:X is OH or OAc;B is

where R₁ is selected from:

-   -   a) R₁₁, where R₁₁ is selected from:        -   1) Hydrogen;        -   2) Deuterium;        -   3) C₁-C₈ alkyl;        -   4) Substituted C₁-C₈ alkyl;        -   5) C₂-C₈ alkenyl;        -   6) Substituted C₂-C₈ alkenyl;        -   7) C₂-C₈ alkynyl;        -   8) Substituted C₂-C₈ alkynyl;        -   9) C₃-C₁₂ cycloalkyl;        -   10) Substituted C₃-C₁₂ cycloalkyl;        -   11) Aryl;        -   12) Substituted aryl;        -   13) Heterocycloalkyl;        -   14) Substituted heterocycloalkyl;        -   15) Heteroaryl; and        -   16) Substituted heteroaryl;    -   b) —C(O)OR₁₁, where R₁₁ is as previously defined;    -   c) —C(O)R₁₁, where R₁₁ is as previously defined;    -   d) —C(O)OCH₂-T-R₁₂, where T is —O— or —S— and R₁₂ is selected        from:        -   1) C₁-C₈ alkyl;        -   2) Substituted C₁-C₈ alkyl;        -   3) C₂-C₈ alkenyl;        -   4) Substituted C₂-C₈ alkenyl;        -   5) C₂-C₈ alkynyl;        -   6) Substituted C₂-C₈ alkynyl;        -   7) C₃-C₁₂ cycloalkyl;        -   8) Substituted C₃-C₁₂ cycloalkyl;        -   9) Aryl;        -   10) Substituted aryl;        -   11) Heterocycloalkyl;        -   12) Substituted heterocycloalkyl;        -   13) Heteroaryl; or        -   14) Substituted heteroaryl;    -   e) —C(O)N(R₁₃)(R₁₄), where R₁₃ and R₁₄ are independently        selected from R₁₁ and R₁₁ is as previously defined or R₁₃ and        R₁₄, together with the nitrogen atom to which they are attached,        form a substituted or unsubstituted heterocycloalkyl;    -   f) —C(O)SR₁₁, where R₁₁ is as previously defined;    -   g) —C(S)OR₁₁, where R₁₁ is as previously defined;    -   h) —C(O)OCH₂OC(O)R₁₂, where R₁₂ is as previously defined;    -   i) —C(S)SR₁₁, where R₁₁ is as previously defined;    -   j) R₁₅, where R₁₅ is selected from:        -   1) -M-R₁₁, where R₁₁ is as previously defined and M is            selected from:            -   i. C₁-C₈ alkyl;            -   ii. Substituted C₁-C₈ alkyl;            -   iii. C₂-C₈ alkenyl;            -   iv. Substituted C₂-C₈ alkenyl;            -   v. C₂-C₈ alkynyl;            -   vi. Substituted C₂-C₈ alkynyl;            -   vii. C₃-C₁₂ cycloalkyl; and            -   viii. Substituted C₃-C₁₂ cycloalkyl;        -   2) -M-NR₁₆R₁₁, where R₁₆ is R₁₁ or R₁₆ and R₁₁ together with            the nitrogen atom to which they are attached form a            substituted or unsubstituted heterocycloalkyl, M is as            previously defined;        -   3) -M-S(O)_(m)R₁₁, where m=0, 1, or 2; M and R₁₁ are as            previously defined;        -   4) -M-OR₁₁, where M and R₁₁ are as previously defined;        -   5) -M-C(O)R₁₁, where M and R₁₁ are as previously defined;        -   6) -M-OC(O)R₁₂, where M and R₁₂ are as previously defined;        -   7) -M-OC(O)OR₁₂, where M and R₁₂ are as previously defined;        -   8) M-NR₁₇C(O)R₁₂, where R₁₇ is R₁₁, M and R₁₂ are as            previously defined;        -   9) -MNR₁₇C(O)OR₁₂, where R₁₇, M and R₁₂ are as previously            defined;        -   10) -M-C(O)NR₁₆R₁₁, where R₁₆, M and R₁₁ are as previously            defined;        -   11) -M-C(O)N(R₁₆)—OR₁₁, where R₁₆, M and R₁₁ are as            previously defined;        -   12) -M-OC(O)NR₁₆R₁₁, where R₁₆, M and R₁₁ are as previously            defined;        -   13) -M-NR₁₇C(O)NR₁₆R₁₁, where M, R₁₁, R₁₇ and R₁₆ are as            previously defined or R₁₆ and R₁₁ together with the nitrogen            atom to which they are attached form a substituted or            unsubstituted heterocycloalkyl;        -   14) -M-C(S)SR₁₁, where M and R₁₁ are as previously defined;        -   15) -M-OC(S)SR₁₂, where M and R₁₂ are as previously defined;        -   16) -M-NR₁₇C(O)SR₁₂, where M, R₁₇ and R₁₂ are as previously            defined;        -   17) -M-SC(O)NR₁₆R₁₁, where M, R₁₁ and R₁₆ are as previously            defined or R₁₆ and R₁₁ together with the nitrogen atom to            which they are attached form a substituted or unsubstituted            heterocycloalkyl;        -   18) -M-CH═N—OR₁₁, where M and R₁₁ are as previously defined;        -   19) -M-CH═N—NR₁₆R₁₁, where M, R₁₁ and R₁₆ are as previously            defined or R₁₆ and R₁₁ together with the nitrogen atom to            which they are attached form a substituted or unsubstituted            heterocycloalkyl;

-   A is ethyl, 1-hydroxyethyl, isopropyl or n-propyl;

-   W and V are each independently absent, —O—, or S(O)_(m)—, where m=0,    1, or 2;

-   R₂ is R₁, where R₁ is as previously defined;

-   R₃ is selected from methyl or ethyl or allyl or propyl;

-   R₄ and R₅ are independently selected from: hydrogen or methyl or    ethyl or allyl, or propyl, or isopropyl;

-   R₆ is R₁, where R₁ is as previously defined; and

-   n and n′ are each independently 0, 1, or 2.

In another embodiment, the present invention provides a pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundor combination of compounds of the present invention, or apharmaceutically acceptable salt form, prodrug, salt of a prodrug,stereoisomer, tautomer, solvate, or combination thereof, in combinationwith a pharmaceutically acceptable carrier or excipient.

In yet another embodiment, the present invention provides a method ofinhibiting the replication of an RNA-containing virus comprisingcontacting said virus with a therapeutically effective amount of acompound or a combination of compounds of the present invention, or apharmaceutically acceptable salt, prodrug, salt of a pro drug,stereoisomer, tautomer, solvate, or combination thereof. Particularly,this invention is directed to methods of inhibiting the replication ofhepatitis C virus.

In still another embodiment, the present invention provides a method oftreating or preventing infection caused by an RNA-containing viruscomprising administering to a patient in need of such treatment atherapeutically effective amount of a compound or combination ofcompounds of the present invention, or a pharmaceutically acceptablesalt form, prodrug, salt of a prodrug, stereoisomer, or tautomer,solvate, or combination thereof. Particularly, this invention isdirected to methods of treating or preventing infection caused byhepatitis C virus.

Yet another embodiment of the present invention provides the use of acompound or combination of compounds of the present invention, or atherapeutically acceptable salt form, prodrug, salt of a prodrug,stereoisomer or tautomer, solvate, or combination thereof, as definedhereinafter, in the preparation of a medicament for the treatment orprevention of infection caused by RNA-containing virus, specificallyhepatitis C virus (HCV).

DETAILED DESCRIPTION OF THE INVENTION

In a first embodiment of the present invention is a compound of formula(I) as illustrated above, or a pharmaceutically acceptable salt, esteror prodrug thereof.

Representative subgenera of the present invention include:

Compounds of formula (I) which are represented by the formula (IIa) or(IIb);

wherein R₁, R₂, R₃, R₄, R₅, R₆, V, and W are as defined above and

represents a single bond or a double bond;

Compounds of formula (I) which are represented by the formula (IIIa) or(IIIb);

wherein R₁, R₂, R₃, R₄, R₅, R₆, V and W are as defined above and

represents a single bond or a double bond.

Compounds of formula (I) which are represented by the formula (IV);

wherein, R₃, R₄, R₅, R₆, and W are as defined above.

Compounds of formula (I) which are represented by the formula (V) or(VI):

wherein R₃, R₄, R₅ and R₆ are as defined above.

Representative species of the present invention include the followingcompounds of formula (V) or (VI):

Example 1 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and R₆=Ac Example2 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and R₆=allyl Example 3Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and R₆=benzyl Example 4Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 5 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 6 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 7 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 8 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 9 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 10 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 11 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 12 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 13 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 14 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 15 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 16 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 17 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 18 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 19 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and R₆=

Example 20 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 21 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 22 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 23 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 24 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 25 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 26 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 27 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 28 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 29 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 30 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 31 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 32 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 33 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 34 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 35 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 36 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 37 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 38 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 39 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 40 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 41 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 42 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 43 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 44 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 45 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 46 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 47 Compound of Formula V: R₃=allyl, R₄═H, R₅═CH₃ and

and

Example 48 Compound of Formula V: R₃═CH₃, R₄═CH₃, R₅═H and R₆=AllylExample 49 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and R₆═H Example50 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and R₆=Allyl Example 51Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and R₆=Benzyl Example 52Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 53 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 54 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 55 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 56 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 57 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 58 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 59 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 60 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 61 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 62 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 63 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 64 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 65 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 66 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 67 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 68 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 69 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and R₆=

Example 70 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 71 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 72 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 73 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 74 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 75 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 76 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 77 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 78 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 79 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 80 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 81 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 82 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 83 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 84 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and R₆=

Example 85 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 86 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 87 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 88 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 89 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 90 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 91 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 92 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 93 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 94 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and

Example 95 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 96 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 97 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 98 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 99 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 100 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 101 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 102 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 103 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 104 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 105 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 106 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 107 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 108 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 109 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 110 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 111 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 112 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 113 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 114 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 115 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 116 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 117 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 118 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 119 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 120 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 121 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 122 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 123 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 124 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 125 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 126 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 127 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 128 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 129 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 130 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

Example 131 Compound of Formula VI: R₃=Me, R₄═H, R₅═CH₃ and

A further embodiment of the present invention includes pharmaceuticalcompositions comprising any single compound delineated herein, or apharmaceutically acceptable salt, ester, solvate, or prodrug thereof,with a pharmaceutically acceptable carrier or excipient.

Yet another embodiment of the present invention is a pharmaceuticalcomposition comprising a combination of two or more compounds delineatedherein, or a pharmaceutically acceptable salt, ester, solvate, orprodrug thereof, with a pharmaceutically acceptable carrier orexcipient.

Yet a further embodiment of the present invention is a pharmaceuticalcomposition comprising any single compound delineated herein incombination with one or more HCV compounds known in the art, or apharmaceutically acceptable salt, ester, solvate, or prodrug thereof,with a pharmaceutically acceptable carrier or excipient.

It will be appreciated that reference herein to therapy and/or treatmentincludes, but is not limited to prevention, retardation, prophylaxis,therapy and cure of the disease. It will further be appreciated thatreferences herein to treatment or prophylaxis of HCV infection includestreatment or prophylaxis of HCV-associated disease such as liverfibrosis, cirrhosis and hepatocellular carcinoma.

It will be further appreciated that the compounds of the presentinvention may contain one or more asymmetric carbon atoms and may existin racemic, diastereoisomeric, and optically active forms. It will stillbe appreciated that certain compounds of the present invention may existin different tautomeric forms. All tautomers are contemplated to bewithin the scope of the present invention.

It will be further appreciated that the compounds of the invention, ortheir pharmaceutically acceptable salts, stereoisomers, tautomers,prodrugs or salt of a prodrug thereof, can be administered as the soleactive pharmaceutical agent, or used in combination with one or moreagents to treat or prevent hepatitis C infections or the symptomsassociated with HCV infection. Other agents to be administered incombination with a compound or combination of compounds of the inventioninclude therapies for disease caused by HCV infection that suppressesHCV viral replication by direct or indirect mechanisms. These includeagents such as host immune modulators (for example, interferon-alpha,pegylated interferon-alpha, interferon-beta, interferon-gamma, CpGoligonucleotides and the like), or antiviral compounds that inhibit hostcellular functions such as inosine monophosphate dehydrogenase (forexample, ribavirin and the like). Also included are cytokines thatmodulate immune function. Also included are vaccines which comprise HCVantigens or antigen adjuvant combinations directed against HCV. Alsoincluded are agents that interact with host cellular components to blockviral protein synthesis by inhibiting the internal ribosome entry site(IRES) initiated translation step of HCV viral replication or to blockviral particle maturation and release with agents targeted toward theviroporin family of membrane proteins such as, for example, HCV P7 andthe like. Other agents to be administered in combination with a compoundof the present invention include any agent or combination of agents thatinhibit the replication of HCV by targeting proteins of the viral genomeinvolved in the viral replication. These agents include but are notlimited to other inhibitors of HCV RNA dependent RNA polymerase such as,for example, nucleoside type polymerase inhibitors described in WO01/90121(A2), or U.S. Pat. No. 6,348,587B1 or WO 01/60315 or WO 01/32153or non-nucleoside inhibitors such as, for example, benzimidazolepolymerase inhibitors described in EP 1 162196A1 or WO 02/04425.

Accordingly, one aspect of the invention is directed to a method fortreating or preventing an infection caused by an RNA-containing viruscomprising co-administering to a patient in need of such treatment oneor more agents selected from the group consisting of a host immunemodulator and a second antiviral agent, or a combination thereof, with atherapeutically effective amount of a compound or combination ofcompounds of the invention, or a pharmaceutically acceptable salt,stereoisomer, tautomer, prodrug, salt of a prodrug, or combinationthereof. Examples of the host immune modulator are, but not limited to,interferon-alpha, pegylated-interferon-alpha, interferon-beta,interferon-gamma, a cytokine, a vaccine, and a vaccine comprising anantigen and an adjuvant, and said second antiviral agent inhibitsreplication of HCV either by inhibiting host cellular functionsassociated with viral replication or by targeting proteins of the viralgenome.

Further aspect of the invention is directed to a method of treating orpreventing infection caused by an RNA-containing virus comprisingco-administering to a patient in need of such treatment an agent orcombination of agents that treat or alleviate symptoms of HCV infectionincluding cirrhosis and inflammation of the liver, with atherapeutically effective amount of a compound or combination ofcompounds of the invention, or a pharmaceutically acceptable salt,stereoisomer, tautomer, prodrug, salt of a prodrug, or combinationthereof. Yet another aspect of the invention provides a method oftreating or preventing infection caused by an RNA-containing viruscomprising co-administering to a patient in need of such treatment oneor more agents that treat patients for disease caused by hepatitis B(HBV) infection, with a therapeutically effective amount of a compoundor a combination of compounds of the invention, or a pharmaceuticallyacceptable salt, stereoisomer, tautomer, prodrug, salt of a prodrug, orcombination thereof. An agent that treats patients for disease caused byhepatitis B (HBV) infection may be for example, but not limited thereto,L-deoxythymidine, adefovir, lamivudine or tenfovir, or any combinationthereof. Example of the RNA-containing virus includes, but not limitedto, hepatitis C virus (HCV).

Another aspect of the invention provides a method of treating orpreventing infection caused by an RNA-containing virus comprisingco-administering to a patient in need of such treatment one or moreagents that treat patients for disease caused by human immunodeficiencyvirus (HIV) infection, with a therapeutically effective amount of acompound or a combination of compounds of the invention, or apharmaceutically acceptable salt, stereoisomer, tautomer, prodrug, saltof a prodrug, or combination thereof. The agent that treats patients fordisease caused by human immunodeficiency virus (HIV) infection mayinclude, but is not limited thereto, ritonavir, lopinavir, indinavir,nelfmavir, saquinavir, amprenavir, atazanavir, tipranavir, TMC-114,fosamprenavir, zidovudine, lamivudine, didanosine, stavudine, tenofovir,zalcitabine, abacavir, efavirenz, nevirapine, delavirdine, TMC-125,L-870812, S-1360, enfuvirtide (T-20) or T-1249, or any combinationthereof. Example of the RNA-containing virus includes, but not limitedto, hepatitis C virus (HCV). In addition, the present invention providesthe use of a compound or a combination of compounds of the invention, ora therapeutically acceptable salt form, stereoisomer, or tautomer,prodrug, salt of a prodrug, or combination thereof, and one or moreagents selected from the group consisting of a host immune modulator anda second antiviral agent, or a combination thereof, to prepare amedicament for the treatment of an infection caused by an RNA-containingvirus in a patient, particularly hepatitis C virus. Examples of the hostimmune modulator are, but not limited to, interferon-alpha,pegylated-interferon-alpha, interferon-beta, interferon-gamma, acytokine, a vaccine, and a vaccine comprising an antigen and anadjuvant, and said second antiviral agent inhibits replication of HCVeither by inhibiting host cellular functions associated with viralreplication or by targeting proteins of the viral genome.

When used in the above or other treatments, combination of compound orcompounds of the invention, together with one or more agents as definedherein above, can be employed in pure form or, where such forms exist,in pharmaceutically acceptable salt form, prodrug, salt of a prodrug, orcombination thereof. Alternatively, such combination of therapeuticagents can be administered as a pharmaceutical composition containing atherapeutically effective amount of the compound or combination ofcompounds of interest, or their pharmaceutically acceptable salt form,prodrugs, or salts of the prodrug, in combination with one or moreagents as defined hereinabove, and a pharmaceutically acceptablecarrier. Such pharmaceutical compositions can be used for inhibiting thereplication of an RNA-containing virus, particularly Hepatitis C virus(HCV), by contacting said virus with said pharmaceutical composition. Inaddition, such compositions are useful for the treatment or preventionof an infection caused by an RNA-containing virus, particularlyHepatitis C virus (HCV).

Hence, further aspect of the invention is directed to a method oftreating or preventing infection caused by an RNA-containing virus,particularly a hepatitis C virus (HCV), comprising administering to apatient in need of such treatment a pharmaceutical compositioncomprising a compound or combination of compounds of the invention or apharmaceutically acceptable salt, stereoisomer, or tautomer, prodrug,salt of a prodrug, or combination thereof, one or more agents as definedhereinabove, and a pharmaceutically acceptable carrier.

When administered as a combination, the therapeutic agents can beformulated as separate compositions which are given at the same time orwithin a predetermined period of time, or the therapeutic agents can begiven as a single unit dosage form.

Antiviral agents contemplated for use in such combination therapyinclude agents (compounds or biologicals) that are effective to inhibitthe formation and/or replication of a virus in a mammal, including butnot limited to agents that interfere with either host or viralmechanisms necessary for the formation and/or replication of a virus ina mammal. Such agents can be selected from another anti-HCV agent; anHIV inhibitor; an HAV inhibitor; and an HBV inhibitor.

Other anti-HCV agents include those agents that are effective fordiminishing or preventing the progression of hepatitis C relatedsymptoms or disease. Such agents include but are not limited toimmunomodulatory agents, inhibitors of HCV NS3 protease, otherinhibitors of HCV polymerase, inhibitors of another target in the HCVlife cycle and other anti-HCV agents, including but not limited toribavirin, amantadine, levovirin and viramidine.

Immunomodulatory agents include those agents (compounds or biologicals)that are effective to enhance or potentiate the immune system responsein a mammal. Immunomodulatory agents include, but are not limited to,inosine monophosphate dehydrogenase inhibitors such as VX-497(merimepodib, Vertex Pharmaceuticals), class I interferons, class IIinterferons, consensus interferons, asialo-interferons pegylatedinterferons and conjugated interferons, including but not limited tointerferons conjugated with other proteins including but not limited tohuman albumin. Class I interferons are a group of interferons that allbind to receptor type I, including both naturally and syntheticallyproduced class I interferons, while class II interferons all bind toreceptor type II. Examples of class I interferons include, but are notlimited to, [alpha]-, [beta]-, [delta]-, [omega]-, and[tau]-interferons, while examples of class II interferons include, butare not limited to, [gamma]-interferons.

Inhibitors of HCV NS3 protease include agents (compounds or biologicals)that are effective to inhibit the function of HCV NS3 protease in amammal Inhibitors of HCV NS3 protease include, but are not limited to,those compounds described in WO 99/07733, WO 99/07734, WO 00/09558, WO00/09543, WO 00/59929, WO 03/064416, WO 03/064455, WO 03/064456, WO2004/030670, WO 2004/037855, WO 2004/039833, WO 2004/101602, WO2004/101605, WO 2004/103996, WO 2005/028501, WO 2005/070955, WO2006/000085, WO 2006/007700 and WO 2006/007708 (all by BoehringerIngelheim), WO 02/060926, WO 03/053349, WO03/099274, WO 03/099316, WO2004/032827, WO 2004/043339, WO 2004/094452, WO 2005/046712, WO2005/051410, WO 2005/054430 (all by BMS), WO 2004/072243, WO2004/093798, WO 2004/113365, WO 2005/010029 (all by Enanta), WO2005/037214 (Intermune) and WO 2005/051980 (Schering), and thecandidates identified as VX-950, ITMN-191 and SCH 503034.

Inhibitors of HCV polymerase include agents (compounds or biologicals)that are effective to inhibit the function of an HCV polymerase. Suchinhibitors include, but are not limited to, non-nucleoside andnucleoside inhibitors of HCV NS5B polymerase. Examples of inhibitors ofHCV polymerase include but are not limited to those compounds describedin: WO 02/04425, WO 03/007945, WO 03/010140, WO 03/010141, WO2004/064925, WO 2004/065367, WO 2005/080388 and WO 2006/007693 (all byBoehringer Ingelheim), WO 2005/049622 (Japan Tobacco), WO 2005/014543(Japan Tobacco) WO 2005/012288 (Genelabs), WO 2004/087714 (IRBM), WO03/101993 (Neogenesis), WO 03/026587 (BMS), WO 03/000254 (JapanTobacco), and WO 01/47883 (Japan Tobacco), and the clinical candidatesXTL-2125, HCV 796, R-1626 and NM 283.

Inhibitors of another target in the HCV life cycle include agents(compounds or biologicals) that are effective to inhibit the formationand/or replication of HCV other than by inhibiting the function of theHCV NS3 protease. Such agents may interfere with either host or HCVviral mechanisms necessary for the formation and/or replication of HCV.Inhibitors of another target in the HCV life cycle include, but are notlimited to, entry inhibitors, agents that inhibit a target selected froma helicase, a NS2/3 protease and an internal ribosome entry site (IRES)and agents that interfere with the function of other viral targetsincluding but not limited to an NS5A protein and an NS4B protein.

It can occur that a patient may be co-infected with hepatitis C virusand one or more other viruses, including but not limited to humanimmunodeficiency virus (HIV), hepatitis A virus (HAV) and hepatitis Bvirus (HBV). Thus also contemplated is combination therapy to treat suchco-infections by co-administering a compound according to the presentinvention with at least one of an HIV inhibitor, an HAV inhibitor and anHBV inhibitor.

Definitions

Listed below are definitions of various terms used to describe thisinvention. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

The term “aryl,” as used herein, refers to a mono-or polycycliccarbocyclic ring system including, but not limited to, phenyl, naphthyl,tetrahydronaphthyl, indanyl, idenyl.

The term “heteroaryl,” as used herein, refers to a mono-or polycyclicaromatic radical having one or more ring atom selected from S, O and N;and the remaining ring atoms are carbon, wherein any N or S containedwithin the ring may be optionally oxidized. Heteroaryl includes, but isnot limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl,imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl,thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzooxazolyl, quinoxalinyl.

In accordance with the invention, any of the aryls, substituted aryls,heteroaryls and substituted heteroaryls described herein, can be anyaromatic group. Aromatic groups can be substituted or unsubstituted.

The terms “C₁-C₈ alkyl,” or “C₁-C₁₂ alkyl,” as used herein, refer tosaturated, straight-or branched-chain hydrocarbon radicals containingbetween one and eight, or one and twelve carbon atoms, respectively.Examples of C₁-C₈ alkyl radicals include, but are not limited to,methyl, ethyl, propyl, isopropyl, n-butyl, tent-butyl, neopentyl,n-hexyl, heptyl and octyl radicals; and examples of C₁-C₁₂ alkylradicals include, but are not limited to, ethyl, propyl, isopropyl,n-hexyl, octyl, decyl, dodecyl radicals.

The term “C₂-C₈ alkenyl,” as used herein, refer to straight-orbranched-chain hydrocarbon radicals containing from two to eight carbonatoms having at least one carbon-carbon double bond by the removal of asingle hydrogen atom. Alkenyl groups include, but are not limited to,for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl,heptenyl, octenyl, and the like.

The term “C₂-C₈ alkynyl,” as used herein, refer to straight-orbranched-chain hydrocarbon radicals containing from two to eight carbonatoms having at least one carbon-carbon triple bond by the removal of asingle hydrogen atom. Representative alkynyl groups include, but are notlimited to, for example, ethynyl, 1-propynyl, 1-butynyl, heptynyl,octynyl, and the like.

The term “C₃-C₈-cycloalkyl”, or “C₃-C₁₂-cycloalkyl,” as used herein,refers to a monocyclic or polycyclic saturated carbocyclic ringcompound. Examples of C₃-C₈-cycloalkyl include, but not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl andcyclooctyl; and examples of C₃-C₁₂-cycloalkyl include, but not limitedto, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,bicyclo[2.2.1]heptyl, and bicyclo[2.2.2]octyl.

The term “C₃-C₈ cycloalkenyl” or “C₃-C₁₂ cycloalkenyl” as used herein,refers to monocyclic or polycyclic carbocyclic ring compound having atleast one carbon-carbon double bond. Examples of C₃-C₈ cycloalkenylinclude, but not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl,cyclohexenyl, cycloheptenyl, cyclooctenyl, and the like; and examples ofC₃-C₁₂ cycloalkenyl include, but not limited to, cyclopropenyl,cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl,and the like.

It is understood that any alkyl, alkenyl, alkynyl and cycloalkyl moietydescribed herein can also be an aliphatic group, an alicyclic group or aheterocyclic group. An “aliphatic” group is a non-aromatic moiety thatmay contain any combination of carbon atoms, hydrogen atoms, halogenatoms, oxygen, nitrogen or other atoms, and optionally contain one ormore units of unsaturation, e.g., double and/or triple bonds. Analiphatic group may be straight chained, branched or cyclic andpreferably contains between about 1 and about 24 carbon atoms, moretypically between about 1 and about 12 carbon atoms. In addition toaliphatic hydrocarbon groups, aliphatic groups include, for example,polyalkoxyalkyls, such as polyalkylene glycols, polyamines, andpolyimines, for example. Such aliphatic groups may be furthersubstituted.

The term “alicyclic,” as used herein, denotes a monovalent group derivedfrom a monocyclic or bicyclic saturated carbocyclic ring compound by theremoval of a single hydrogen atom. Examples include, but not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[2.2.1]heptyl,and bicyclo[2.2.2]octyl. Such alicyclic groups may be furthersubstituted.

The terms “heterocyclic” or “heterocycloalkyl” can be usedinterchangeably and referred to a non-aromatic ring or a bi-ortri-cyclic group fused system, where (i) each ring system contains atleast one heteroatom independently selected from oxygen, sulfur andnitrogen, (ii) each ring system can be saturated or unsaturated (iii)the nitrogen and sulfur heteroatoms may optionally be oxidized, (iv) thenitrogen heteroatom may optionally be quaternized, (v) any of the aboverings may be fused to an aromatic ring, and (vi) the remaining ringatoms are carbon atoms which may be optionally oxo-substituted.Representative heterocyclic groups include, but are not limited to,1,3-dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl,imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl,morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl,pyridazinonyl, and tetrahydrofuryl. Such heterocyclic groups may befurther substituted.

The term “substituted” refers to substitution by independent replacementof one, two, or three or more of the hydrogen atoms thereon withsubstituents including, but not limited to, —F, —Cl, —Br, —I, —OH,protected hydroxy, —NO₂, —CN, —N₃, —NH₂, protected amino, oxo, thioxo,—NH—C₁-C₁₂-alkyl, —NH—C₂-C₈-alkenyl, —NH—C₂-C₈-alkynyl,—NH—C₃-C₁₂-cycloalkyl, —NH-aryl, —NH-heteroaryl, —NH-heterocycloalkyl,-dialkylamino, -diarylamino, -diheteroarylamino, —O—C₁-C₁₂-alkyl,—O—C₂-C₈-alkenyl, —O—C₂-C₈-alkynyl, —O—C₃-C₁₂-cycloalkyl, —O-aryl,—O-heteroaryl, —O-heterocycloalkyl, —C(O)—C₁-C₁₂-alkyl,—C(O)—C₂-C₈-alkenyl, —C(O)—C₂-C₈-alkynyl, —C(O)—C₃-C₁₂-cycloalkyl,—C(O)-aryl, —C(O)-heteroaryl, —C(O)-heterocycloalkyl, —CONH₂,—CONH—C₁-C₁₂-alkyl, —CONH—C₂-C₈-alkenyl, —CONH—C₂-C₈-alkynyl,—CONH—C₃-C₁₂-cycloalkyl, —CONH-aryl, —CONH-heteroaryl,—CONH-heterocycloalkyl, —OCO₂—C₁-C₁₂-alkyl, —OCO₂—C₂-C₈-alkenyl,—OCO₂—C₂-C₈-alkynyl, —OCO₂—C₃-C₁₂-cycloalkyl, —OCO₂-aryl,—OCO₂-heteroaryl, —OCO₂-heterocycloalkyl, —OCONH₂, —OCONH—C₁-C₁₂-alkyl,—OCONH—C₂-C₈-alkenyl, —OCONH—C₂-C₈-alkynyl, —OCONH—C₃-C₁₂-cycloalkyl,—OCONH-aryl, —OCONH-heteroaryl, —OCONH— heterocycloalkyl,—NHC(O)—C₁-C₁₂-alkyl, —NHC(O)—C₂-C₈-alkenyl, —NHC(O)—C₂-C₈-alkynyl,—NHC(O)—C₃-C₁₂-cycloalkyl, —NHC(O)-aryl, —NHC(O)-heteroaryl,—NHC(O)-heterocycloalkyl, —NHCO₂—C₁-C₁₂-alkyl, —NHCO₂—C₂-C₈-alkenyl,—NHCO₂—C₂-C₈-alkynyl, —NHCO₂—C₃-C₁₂-cycloalkyl, —NHCO₂-aryl,—NHCO₂-heteroaryl, —NHCO₂— heterocycloalkyl, —NHC(O)NH₂,—NHC(O)NH—C₁-C₁₂-alkyl, —NHC(O)NH—C₂-C₈-alkenyl,—NHC(O)NH—C₂-C₈-alkynyl, —NHC(O)NH—C₃-C₁₂-cycloalkyl, —NHC(O)NH-aryl,—NHC(O)NH-heteroaryl, —NHC(O)NH-heterocycloalkyl, NHC(S)NH₂,—NHC(S)NH—C₁-C₁₂-alkyl, —NHC(S)NH—C₂-C₈-alkenyl,—NHC(S)NH—C₂-C₈-alkynyl, —NHC(S)NH—C₃-C₁₂-cycloalkyl, —NHC(S)NH-aryl,—NHC(S)NH-heteroaryl, —NHC(S)NH-heterocycloalkyl, —NHC(NH)NH₂,—NHC(NH)NH—C₁-C₁₂-alkyl, —NHC(NH)NH—C₂-C₈-alkenyl,—NHC(NH)NH—C₂-C₈-alkynyl, —NHC(NH)NH—C₃-C₁₂-cycloalkyl, —NHC(NH)NH-aryl,—NHC(NH)NH-heteroaryl, —NHC(NH)NH-heterocycloalkyl,—NHC(NH)—C₁-C₁₂-alkyl, —NHC(NH)—C₂-C₈-alkenyl, —NHC(NH)—C₂-C₈-alkynyl,—NHC(NH)—C₃-C₁₂-cycloalkyl, —NHC(NH)-aryl, —NHC(NH)-heteroaryl,—NHC(NH)-heterocycloalkyl, —C(NH)NH—C₁-C₁₂-alkyl,—C(NH)NH—C₂-C₈-alkenyl, —C(NH)NH—C₂-C₈-alkynyl,—C(NH)NH—C₃-C₁₂-cycloalkyl, —C(NH)NH-aryl, —C(NH)NH-heteroaryl,—C(NH)NH-heterocycloalkyl, —S(O)—C₁-C₁₂-alkyl, —S(O)—C₂-C₈-alkenyl,—S(O)—C₂-C₈-alkynyl, —S(O)—C₃-C₁₂-cycloalkyl, —S(O)-aryl,—S(O)-heteroaryl, —S(O)-heterocycloalkyl, —SO₂NH₂, —SO₂NH—C₁-C₁₂-alkyl,—SO₂NH—C₂-C₈-alkenyl, —SO₂NH—C₂-C₈-alkynyl, —SO₂NH—C₃-C₁₂-cycloalkyl,—SO₂NH-aryl, —SO₂NH-heteroaryl, —SO₂NH-heterocycloalkyl,—NHSO₂—C₁-C₁₂-alkyl, —NHSO₂—C₂-C₈-alkenyl, —NHSO₂—C₂-C₈-alkynyl,—NHSO₂—C₃-C₁₂-cycloalkyl, —NHSO₂-aryl, —NHSO₂-heteroaryl,—NHSO₂-heterocycloalkyl, —CH₂NH₂, —CH₂SO₂CH₃, -aryl, -arylalkyl,-heteroaryl, -heteroarylalkyl, -heterocycloalkyl, —C₃-C₁₂-cycloalkyl,polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, —SH,—S—C₁-C₁₂-alkyl, —S—C₂-C₈-alkenyl, —S—C₂-C₈-alkynyl,—S—C₃-C₁₂-cycloalkyl, —S-aryl, —S-heteroaryl, —S-heterocycloalkyl, ormethylthiomethyl. It is understood that the aryls, heteroaryls, alkyls,and the like can be further substituted.

The term “halogen,” as used herein, refers to an atom selected fromfluorine, chlorine, bromine and iodine.

The term “hydroxy activating group”, as used herein, refers to a labilechemical moiety which is known in the art to activate a hydroxyl groupso that it will depart during synthetic procedures such as in asubstitution or an elimination reaction. Examples of hydroxyl activatinggroup include, but not limited to, mesylate, tosylate, triflate,p-nitrobenzoate, phosphonate and the like.

The term “activated hydroxy”, as used herein, refers to a hydroxy groupactivated with a hydroxyl activating group, as defined above, includingmesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups, forexample.

The term “hydroxy protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect a hydroxyl groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the hydroxy protecting group as described hereinmay be selectively removed. Hydroxy protecting groups as known in theart are described generally in T. H. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons,New York (1999). Examples of hydroxyl protecting groups includebenzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 4-bromobenzyloxycarbonyl,4-methoxybenzyloxycarbonyl, methoxycarbonyl, tert-butoxycarbonyl,isopropoxycarbonyl, diphenylmethoxycarbonyl,2,2,2-trichloroethoxycarbonyl, 2-(trimethylsilyl)ethoxycarbonyl,2-furfuryloxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl,trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, t-butyl,2,2,2-trichloroethyl, 2-trimethylsilyl ethyl, 1,1-dimethyl-2-propenyl,3-methyl-3-butenyl, allyl, benzyl, para-methoxybenzyldiphenylmethyl,triphenylmethyl (trityl), tetrahydrofuryl, methoxymethyl,methylthiomethyl, benzyloxymethyl, 2,2,2-triehloroethoxymethyl,2-(trimethylsilyl)ethoxymethyl, methanesulfonyl, para-toluenesulfonyl,trimethylsilyl, triethylsilyl, triisopropylsilyl, and the like.Preferred hydroxyl protecting groups for the present invention areacetyl (Ac or —C(O)CH₃), benzoyl (Bz or —C(O)C₆H₅), and trimethylsilyl(TMS or —Si(CH₃)₃).

The term “protected hydroxy,” as used herein, refers to a hydroxy groupprotected with a hydroxy protecting group, as defined above, includingbenzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl groups,for example.

The term “hydroxy prodrug group”, as used herein, refers to a promoietygroup which is known in the art to change the physicochemical, and hencethe biological properties of a parent drug in a transient manner bycovering or masking the hydroxy group. After said syntheticprocedure(s), the hydroxy prodrug group as described herein must becapable of reverting back to hydroxy group in vivo. Hydroxy prodruggroups as known in the art are described generally in Kenneth B. Sloan,Prodrugs, Topical and Ocular Drug Delivery, (Drugs and thePharmaceutical Sciences; Volume 53), Marcel Dekker, Inc., New York(1992).

The term “amino protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect an amino groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the amino protecting group as described hereinmay be selectively removed. Amino protecting groups as known in the artare described generally in T. H. Greene and P. G. M. Wuts, ProtectiveGroups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York(1999). Examples of amino protecting groups include, but are not limitedto, t-butoxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyloxycarbonyl, andthe like.

The term “leaving group” means a functional group or atom which can bedisplaced by another functional group or atom in a substitutionreaction, such as a nucleophilic substitution reaction. By way ofexample, representative leaving groups include chloro, bromo and iodogroups; sulfonic ester groups, such as mesylate, tosylate, brosylate,nosylate and the like; and acyloxy groups, such as acetoxy,trifluoroacetoxy and the like.

The term “protected amino,” as used herein, refers to an amino groupprotected with an amino protecting group as defined above.

The term “aprotic solvent,” as used herein, refers to a solvent that isrelatively inert to proton activity, i.e., not acting as a proton-donor.Examples include, but are not limited to, hydrocarbons, such as hexaneand toluene, for example, halogenated hydrocarbons, such as, forexample, methylene chloride, ethylene chloride, chloroform, and thelike, heterocyclic compounds, such as, for example, tetrahydrofuran andN-methylpyrrolidinone, and ethers such as diethyl ether,bis-methoxymethyl ether. Such compounds are well known to those skilledin the art, and it will be obvious to those skilled in the art thatindividual solvents or mixtures thereof may be preferred for specificcompounds and reaction conditions, depending upon such factors as thesolubility of reagents, reactivity of reagents and preferred temperatureranges, for example. Further discussions of aprotic solvents may befound in organic chemistry textbooks or in specialized monographs, forexample: Organic Solvents Physical Properties and Methods ofPurification, 4th ed., edited by John A. Riddick et al., Vol. II, in theTechniques of Chemistry Series, John Wiley & Sons, NY, 1986.

The term “protic solvent’ as used herein, refers to a solvent that tendsto provide protons, such as an alcohol, for example, methanol, ethanol,propanol, isopropanol, butanol, t-butanol, and the like. Such solventsare well known to those skilled in the art, and it will be obvious tothose skilled in the art that individual solvents or mixtures thereofmay be preferred for specific compounds and reaction conditions,depending upon such factors as the solubility of reagents, reactivity ofreagents and preferred temperature ranges, for example. Furtherdiscussions of protogenic solvents may be found in organic chemistrytextbooks or in specialized monographs, for example: Organic SolventsPhysical Properties and Methods of Purification, 4th ed., edited by JohnA. Riddick et al., Vol. II, in the Techniques of Chemistry Series, JohnWiley & Sons, NY, 1986.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable”, as used herein, refers to compounds which possessstability sufficient to allow manufacture and which maintains theintegrity of the compound for a sufficient period of time to be usefulfor the purposes detailed herein (e.g., therapeutic or prophylacticadministration to a subject).

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography, highpressure liquid chromatography, or recrystallization. As can beappreciated by the skilled artisan, further methods of synthesizing thecompounds of the Formula herein will be evident to those of ordinaryskill in the art. Additionally, the various synthetic steps may beperformed in an alternate sequence or order to give the desiredcompounds. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing thecompounds described herein are known in the art and include, forexample, those such as described in R. Larock, Comprehensive OrganicTransformations, 2^(nd) Ed. Wiley-VCH (1999); T. W. Greene and P. G. M.Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley andSons (1999); L. Fieser and M. Fieser, Fieser and Fieser's Reagents forOrganic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons(1995), and subsequent editions thereof.

The term “subject” as used herein refers to an animal. Preferably theanimal is a mammal. More preferably the mammal is a human. A subjectalso refers to, for example, dogs, cats, horses, cows, pigs, guineapigs, fish, birds and the like.

The compounds of this invention may be modified by appending appropriatefunctionalities to enhance selective biological properties. Suchmodifications are known in the art and may include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of excretion.

The compounds described herein contain one or more asymmetric centersand thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)-or (S)-, or as (D)-or (L)-for amino acids. Thepresent invention is meant to include all such possible isomers, as wellas their racemic and optically pure forms. Optical isomers may beprepared from their respective optically active precursors by theprocedures described above, or by resolving the racemic mixtures. Theresolution can be carried out in the presence of a resolving agent, bychromatography or by repeated crystallization or by some combination ofthese techniques which are known to those skilled in the art. Furtherdetails regarding resolutions can be found in Jacques, et al.,Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). Whenthe compounds described herein contain olefinic double bonds, otherunsaturation, or other centers of geometric asymmetry, and unlessspecified otherwise, it is intended that the compounds include both Eand Z geometric isomers or cis-and trans-isomers. Likewise, alltautomeric forms are also intended to be included. Tautomers may be incyclic or acyclic. The configuration of any carbon-carbon double bondappearing herein is selected for convenience only and is not intended todesignate a particular configuration unless the text so states; thus acarbon-carbon double bond or carbon-heteroatom double bond depictedarbitrarily herein as trans may be cis, trans, or a mixture of the twoin any proportion.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge, etal. describes pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 66: 1-19 (1977). The salts can be prepared insitu during the final isolation and purification of the compounds of theinvention, or separately by reacting the free base function with asuitable organic acid. Examples of pharmaceutically acceptable saltsinclude, but are not limited to, nontoxic acid addition salts are saltsof an amino group formed with inorganic acids such as hydrochloric acid,hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid orwith organic acids such as acetic acid, maleic acid, tartaric acid,citric acid, succinic acid or malonic acid or by using other methodsused in the art such as ion exchange. Other pharmaceutically acceptablesalts include, but are not limited to, adipate, alginate, ascorbate,aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate,hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like.Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and arylsulfonate.

As used herein, the term “pharmaceutically acceptable ester” refers toesters which hydrolyze in vivo and include those that break down readilyin the human body to leave the parent compound or a salt thereof.Suitable ester groups include, for example, those derived frompharmaceutically acceptable aliphatic carboxylic acids, particularlyalkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which eachalkyl or alkenyl moiety advantageously has not more than 6 carbon atoms.Examples of particular esters include, but are not limited to, formates,acetates, propionates, butyrates, acrylates and ethylsuccinates.

The term “pharmaceutically acceptable prodrugs” as used herein refers tothose prodrugs of the compounds of the present invention which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and lower animals with undue toxicity,irritation, allergic response, and the like, commensurate with areasonable benefit/risk ratio, and effective for their intended use, aswell as the zwitterionic forms, where possible, of the compounds of thepresent invention. “Prodrug”, as used herein means a compound which isconvertible in vivo by metabolic means (e.g. by hydrolysis) to acompound of the invention. Various forms of prodrugs are known in theart, for example, as discussed in Bundgaard, (ed.), Design of Prodrugs,Elsevier (1985); Widder, et al. (ed.), Methods in Enzymology, vol. 4,Academic Press (1985); Krogsgaard-Larsen, et al., (ed). “Design andApplication of Prodrugs, Textbook of Drug Design and Development,Chapter 5, 113-191 (1991); Bundgaard, et al., Journal of Drug DeliverReviews, 8:1-38 (1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285et seq. (1988); Higuchi and Stella (eds.) Prodrugs as Novel DrugDelivery Systems, American Chemical Society (1975); and Bernard Testa &Joachim Mayer, “Hydrolysis In Drug And Prodrug Metabolism: Chemistry,Biochemistry And Enzymology,” John Wiley and Sons, Ltd. (2002).

The present invention also relates to solvates of the compounds ofFormula (I), for example hydrates.

This invention also encompasses pharmaceutical compositions containing,and methods of treating viral infections through administering,pharmaceutically acceptable prodrugs of compounds of the invention. Forexample, compounds of the invention having free amino, amido, hydroxy orcarboxylic groups can be converted into prodrugs. Prodrugs includecompounds wherein an amino acid residue, or a polypeptide chain of twoor more (e.g., two, three or four) amino acid residues is covalentlyjoined through an amide or ester bond to a free amino, hydroxy orcarboxylic acid group of compounds of the invention. The amino acidresidues include but are not limited to the 20 naturally occurring aminoacids commonly designated by three letter symbols and also includes4-hydroxyproline, hydroxylysine, demosine, isodemosine,3-methylhistidine, norvalin, beta-alanine, gamma-PAGE aminobutyric acid,citrulline, homocysteine, homoserine, ornithine and methionine sulfone.Additional types of prodrugs are also encompassed. For instance, freecarboxyl groups can be derivatized as amides or alkyl esters. Freehydroxy groups may be derivatized using groups including but not limitedto hemisuccinates, phosphate esters, dimethylaminoacetates, andphosphoryloxymethyloxycarbonyls, as outlined in Advanced Drug DeliveryReviews, 1996, 19, 115. Carbamate prodrugs of hydroxy and amino groupsare also included, as are carbonate prodrugs, sulfonate esters andsulfate esters of hydroxy groups. Derivatization of hydroxy groups as(acyloxy)methyl and (acyloxy)ethyl ethers wherein the acyl group may bean alkyl ester, optionally substituted with groups including but notlimited to ether, amine and carboxylic acid functionalities, or wherethe acyl group is an amino acid ester as described above, are alsoencompassed. Prodrugs of this type are described in J. Med. Chem. 1996,39, 10. Free amines can also be derivatized as amides, sulfonamides orphosphonamides. All of these prodrug moieties may incorporate groupsincluding but not limited to ether, amine and carboxylic acidfunctionalities.

Pharmaceutical Compositions

The pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the present inventionformulated together with one or more pharmaceutically acceptablecarriers or excipients.

As used herein, the term “pharmaceutically acceptable carrier orexcipient” means a non-toxic, inert solid, semi-solid or liquid filler,diluent, encapsulating material or formulation auxiliary of any type.Some examples of materials which can serve as pharmaceuticallyacceptable carriers are sugars such as lactose, glucose and sucrose;starches such as corn starch and potato starch; cellulose and itsderivatives such as sodium carboxymethyl cellulose, ethyl cellulose andcellulose acetate; powdered tragacanth; malt; gelatin; talc; excipientssuch as cocoa butter and suppository waxes; oils such as peanut oil,cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; glycols such as propylene glycol; esters such as ethyloleate and ethyl laurate; agar; buffering agents such as magnesiumhydroxide and aluminum hydroxide; alginic acid; pyrogen-free water;isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffersolutions, as well as other non-toxic compatible lubricants such assodium lauryl sulfate and magnesium stearate, as well as coloringagents, releasing agents, coating agents, sweetening, flavoring andperfuming agents, preservatives and antioxidants can also be present inthe composition, according to the judgment of the formulator.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir, preferably by oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. In somecases, the pH of the formulation may be adjusted with pharmaceuticallyacceptable acids, bases or buffers to enhance the stability of theformulated compound or its delivery form. The term parenteral as usedherein includes subcutaneous, intracutaneous, intravenous,intramuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions, may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono-or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolution,which, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions that are compatible with body tissues. Compositions forrectal or vaginal administration are preferably suppositories which canbe prepared by mixing the compounds of this invention with suitablenon-irritating excipients or carriers such as cocoa butter, polyethyleneglycol or a suppository wax which are solid at ambient temperature butliquid at body temperature and therefore melt in the rectum or vaginalcavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or: a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, eye ointments, powders and solutionsare also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

For pulmonary delivery, a therapeutic composition of the invention isformulated and administered to the patient in solid or liquidparticulate form by direct administration e.g., inhalation into therespiratory system. Solid or liquid particulate forms of the activecompound prepared for practicing the present invention include particlesof respirable size: that is, particles of a size sufficiently small topass through the mouth and larynx upon inhalation and into the bronchiand alveoli of the lungs. Delivery of aerosolized therapeutics,particularly aerosolized antibiotics, is known in the art (see, forexample U.S. Pat. No. 5,767,068 to VanDevanter et al., U.S. Pat. No.5,508,269 to Smith et al., and WO 98/43650 by Montgomery, all of whichare incorporated herein by reference). A discussion of pulmonarydelivery of antibiotics is also found in U.S. Pat. No. 6,014,969,incorporated herein by reference.

According to the methods of treatment of the present invention, viralinfections, conditions are treated or prevented in a patient such as ahuman or another animal by administering to the patient atherapeutically effective amount of a compound of the invention, in suchamounts and for such time as is necessary to achieve the desired result.

By a “therapeutically effective amount” of a compound of the inventionis meant an amount of the compound which confers a therapeutic effect onthe treated subject, at a reasonable benefit/risk ratio applicable toany medical treatment. The therapeutic effect may be objective (i.e.,measurable by some test or marker) or subjective (i.e., subject gives anindication of or feels an effect). An effective amount of the compounddescribed above may range from about 0.1 mg/Kg to about 500 mg/Kg,preferably from about 1 to about 50 mg/Kg. Effective doses will alsovary depending on route of administration, as well as the possibility ofco-usage with other agents. It will be understood, however, that thetotal daily usage of the compounds and compositions of the presentinvention will be decided by the attending physician within the scope ofsound medical judgment. The specific therapeutically effective doselevel for any particular patient will depend upon a variety of factorsincluding the disorder being treated and the severity of the disorder;the activity of the specific compound employed; the specific compositionemployed; the age, body weight, general health, sex and diet of thepatient; the time of administration, route of administration, and rateof excretion of the specific compound employed; the duration of thetreatment; drugs used in combination or contemporaneously with thespecific compound employed; and like factors well known in the medicalarts.

The total daily dose of the compounds of this invention administered toa human or other animal in single or in divided doses can be in amounts,for example, from 0.01 to 50 mg/kg body weight or more usually from 0.1to 25 mg/kg body weight. Single dose compositions may contain suchamounts or submultiples thereof to make up the daily dose. In general,treatment regimens according to the present invention compriseadministration to a patient in need of such treatment from about 10 mgto about 1000 mg of the compound(s) of this invention per day in singleor multiple doses.

The compounds of the Formula described herein can, for example, beadministered by injection, intravenously, intraarterially, subdermally,intraperitoneally, intramuscularly, or subcutaneously; or orally,buccally, nasally, transmucosally, topically, in an ophthalmicpreparation, or by inhalation, with a dosage ranging from about 0.1 toabout 500 mg/kg of body weight, alternatively dosages between 1 mg and1000 mg/dose, every 4 to 120 hours, or according to the requirements ofthe particular drug. The methods herein contemplate administration of aneffective amount of compound or compound composition to achieve thedesired or stated effect. Typically, the pharmaceutical compositions ofthis invention will be administered from about 1 to about 6 times perday or alternatively, as a continuous infusion. Such administration canbe used as a chronic or acute therapy. The amount of active ingredientthat may be combined with pharmaceutically exipients or carriers toproduce a single dosage form will vary depending upon the host treatedand the particular mode of administration. A typical preparation willcontain from about 5% to about 95% active compound (w/w). Alternatively,such preparations may contain from about 20% to about 80% activecompound.

Lower or higher doses than those recited above may be required. Specificdosage and treatment regimens for any particular patient will dependupon a variety of factors, including the activity of the specificcompound employed, the age, body weight, general health status, sex,diet, time of administration, rate of excretion, drug combination, theseverity and course of the disease, condition or symptoms, the patient'sdisposition to the disease, condition or symptoms, and the judgment ofthe treating physician.

Upon improvement of a patient's condition, a maintenance dose of acompound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level. Patients may, however,require intermittent treatment on a long-term basis upon any recurrenceof disease symptoms.

When the compositions of this invention comprise a combination of acompound of the invention described herein and one or more additionaltherapeutic or prophylactic agents, both the compound and the additionalagent should be present at dosage levels of between about 1 to 100%, andmore preferably between about 5 to 95% of the dosage normallyadministered in a monotherapy regimen. The additional agents may beadministered separately, as part of a multiple dose regimen, from thecompounds of this invention. Alternatively, those agents may be part ofa single dosage form, mixed together with the compounds of thisinvention in a single composition.

The said “additional therapeutic or prophylactic agents” includes butnot limited to, immune therapies (eg. interferon), therapeutic vaccines,antifibrotic agents, anti-inflammatory agents such as corticosteroids orNSAIDs, bronchodilators such as beta-2 adrenergic agonists and xanthines(e.g. theophylline), mucolytic agents, anti-muscarinics,anti-leukotrienes, inhibitors of cell adhesion (e.g. ICAM antagonists),anti-oxidants (eg N-acetylcysteine), cytokine agonists, cytokineantagonists, lung surfactants and/or antimicrobial and anti-viral agents(eg ribavirin and amantidine). The compositions according to theinvention may also be used in combination with gene replacement therapy.

Unless otherwise defined, all technical and scientific terms used hereinare accorded the meaning commonly known to one of ordinary skill in theart. All publications, patents, published patent applications, and otherreferences mentioned herein are hereby incorporated by reference intheir entirety.

Abbreviations

Abbreviations which may be used in the descriptions of the scheme andthe examples that follow are:

-   -   Ac for acetyl;    -   Boc₂O for di-tert-butyl-dicarbonate;    -   Boc for t-butoxycarbonyl;    -   Bz for benzoyl;    -   Bn for benzyl;    -   BocNHOH for tent-butyl N-hydroxycarbamate;    -   t-BuOK for potassium tert-butoxide;    -   BOP for (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium        Hexafluorophosphate;    -   Brine for sodium chloride solution in water;    -   CDI for carbonyldiimidazole;    -   CH₂Cl₂ for dichloromethane;    -   CH₃ for methyl;    -   CH₃CN for acetonitrile;    -   Cs₂CO₃ for cesium carbonate;    -   dba for dibenzylidene acetone;    -   dppb for diphenylphosphino butane;    -   dppe for diphenylphosphino ethane;    -   DBU for 1,8-diazabicyclo[5.4.0]undec-7-ene;    -   DCC for N,N′-dicyclohexylcarbodiimide;    -   DEAD for diethylazodicarboxylate;    -   DIAD for diisopropyl azodicarboxylate;    -   DIPEA or (i-Pr)₂EtN for N,N,-diisopropylethyl amine;    -   Dess-Martin periodinane for        1,1,1-tris(acetyloxy)-1,1-dihydro-1,2-benziodoxol-3-(1H)-one;    -   DMAP for 4-dimethylaminopyridine;    -   DME for 1,2-dimethoxyethane;    -   DMF for N,N-dimethylformamide;    -   DMSO for dimethyl sulfoxide;    -   DPPA for diphenylphosphoryl azide;    -   EDC for N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide;    -   EDC HCl for N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide        hydrochloride; EtOAc for ethyl acetate;    -   EtOH for ethanol;    -   Et₂O for diethyl ether;    -   HATU for        O-(7-azabenzotriazol-1-yl)-N,N,N′,N′,-tetramethyluronium        Hexafluorophosphate;    -   HCl for hydrogen chloride;    -   HOBT for 1-hydroxybenzotriazole;    -   K₂CO₃ for potassium carbonate;    -   MeOH for methanol;    -   Ms for mesyl or —SO₂—CH₃;    -   Ms₂O for methanesulfonic anhydride or mesyl-anhydride;    -   NaHCO₃ for sodium bicarbonate or sodium hydrogen carbonate;    -   Na₂CO₃ sodium carbonate;    -   NaOH for sodium hydroxide;    -   Na₂SO₄ for sodium sulfate;    -   NaHSO₃ for sodium bisulfite or sodium hydrogen sulfite;    -   Na₂S₂O₃ for sodium thiosulfate;    -   NH₂NH₂ for hydrazine;    -   NH₄HCO₃ for ammonium bicarbonate;    -   NH₄Cl for ammonium chloride;    -   NMMO for N-methylmorpholine N-oxide;    -   NaIO₄ for sodium periodate;    -   OH for hydroxy;    -   OsO₄ for osmium tetroxide;    -   TEA or Et₃N for triethylamine;    -   TFA for trifluoroacetic acid;    -   THF for tetrahydrofuran;    -   TPP or PPh₃ for triphenylphosphine;    -   Ts for tosyl or SO₂—C₆H₄CH₃;    -   Ts₂O for tolylsulfonic anhydride or tosyl-anhydride;    -   TsOH for p-tolylsulfonic acid;    -   Pd for palladium;    -   Ph for phenyl;    -   Pd₂(dba)₃ for tris(dibenzylideneacetone) dipalladium (0);    -   Pd(PPh₃)₄ for tetrakis(triphenylphosphine)palladium (0);    -   TBS for tent-butyl dimethylsilyl; or    -   TMS for trimethylsilyl;    -   TMSCl for trimethylsilyl chloride; and    -   CsA for cyclosporin A.        Synthetic Methods

The compounds and processes of the present invention will be betterunderstood in connection with the following synthetic schemes thatillustrate the methods by which the compounds of the invention may beprepared.

The novel cyclosporin analogues of the present invention are derivedfrom cyclosporin A. As shown in Scheme 1, a key intermediate of formula(1-3) was prepared by selective removal of amino acid in positionfour—N-methyl leucine of cyclosporin A (see Roland Wenger et al,“Synthetic routes to NEtXaa⁴-cycloporin A derivatives as potentialanti-HIV I drugs”, Tetrahedron Letters, 2000, 41, 7193, which is herebyincorporated by reference in its entirety). Thus, cyclosporin A wasreacted with acetic anhydride, optionally in the presence of pyridine orDMAP in CH₂Cl₂ to give acetylated intermediate (1-1), which was followedby selective cleavage of the amide bond between position three andposition four amino acid with trimethyloxonium tetrafluoroborate inCH₂Cl₂ to afford the intermediate (1-2). Edman degradation of (1-2) gavethe key intermediate (1-3).

Scheme 2 illustrates a process of the invention for the preparation ofcompounds according to the invention. The intermediate of formula (1-3),is then converted to the compound of formula (2-1) by hydrolysispromoted with inorganic base such as but not limited to sodiummethoxide, potassium carbonate, sodium carbonate, and the like. Thereaction is carried our in a solvent such as but not limited tomethanol, ethanol, THF, DMF, CH₃CN. The most preferred solvent ismethanol. The reaction temperature can vary from 0° C. to about 50° C.

Then the compound of formula (2-1) is coupled with a protected aminoacid of the formula (2-2), where R₆, R₃, R₄ and R₅ are as previouslydefined to give the compound of formula (2-3). The coupling regent canbe selected from, but not limited to DCC, EDC, di-isopropylcarbodiimide, BOP-Cl, PyBOP, PyAOP, TFFH and HATU. Suitable basesinclude, but are not limited to, triethylamine, diisopropylethylamine,DBU, N-methylmorpholine and DMAP. The coupling reaction is carried outin an aprotic solvent such as, but not limited to, CH₂Cl₂, DMF and THF.The reaction temperature can vary from 0° C. to about 50° C.

The protected amino acids of formula (2-2) are prepared by the methoddescribed in Hu, T. and Panek, J. S.; J. Am. Chem. Soc. 2002, 124,11372.

The methyl ester of compound of formula (2-3) is converted to thecorresponding acid compound of formula (2-4) via alkaline hydrolysis inprotic solvents. Representative alkali compounds include lithiumhydroxide, sodium hydroxide, potassium hydroxide, and the like. Suitablesolvents include, but are not limited to, methanol, ethanol,isopropanol, butanol, THF, 1,4-dioxane and mixtures there of. Thereaction temperature is preferably 0° to 35° C.

Compound of formula (2-4) is converted to the compound of formula (2-5)by acidic Boc deprotection. The acid can be selected from, but notlimited to, TFA, HCl in dioxane, methanesulfonic acid. A more throughdiscussion of the precedures, reagents and conditions for removingprotecting groups is described in literature, for example, by T. W.Greene and P. G. M. Wuts in “Pretective Groups in Organic Synthesis”3^(rd) ed., John Wiley & Son, Inc., 1999.

Compound of formula (2-6) is prepared by intramolecular amide formationreaction. The regent can be selected from, but not limited to DCC, EDC,di-isopropyl carbodiimide, BOP-Cl, PyBOP, PyAOP, TFFH and HATU. Suitablebases include, but are not limited to, triethylamine,diisopropylethylamine, DBU, N-methylmorpholine and DMAP. The couplingreaction is carried out in an aprotic solvent such as, but not limitedto, CH₂Cl₂, DMF and THF. The reaction temperature can vary from 0° C. toabout 50° C.

An alternative process for the preparation of the novel cyclosporinanalogues of the present invention is also illustrated in Scheme 3.

The amino group of compound of formula (2-1) is protected by reactingwith Boc₂O in the presence of a base such as, but limited to TEA, DIPEA,DMAP, pyridine and the like. The reaction can be carried out in avariety of organic solvents such as CH₂Cl₂, toluene, Et₂O, EtOAc andchloroform. Then the hydroxyl group is protected with silylatingreagents such as, but not limited to, TMSCl, TBSCl, TESCl, TMSOTf andN,O-bis(trimethylsilyl)acetamide in the presence of an organic base.Preferably, the silylating reagent is TMSCl andN,O-bis-(trimethylsilyl)acetamide, the organic base is1-methylimidazole. A more through discussion of the procedures, reagentsand conditions for protecting hydroxyl group is described in literature,for example, by T. W. Greene and P. G. M. Wuts in “Pretective Groups inOrganic Synthesis” 3^(rd) ed., John Wiley & Son, Inc., 1999.

The compound of formula (3-1) is converted to the compound of formula(3-2) by hydrolysis with the essential same condition described in theconversion of (1-3) to (2-1). Further coupling of (3-2) with a protectedamino acid of formula (3-3) is proceeded using essential same conditiondescribed in the conversion of (2-1) to (2-3) to give the compound offormula (3-4).

Compound of formula (3-4) is converted to the compound of formula (3-5)by acidic N-Boc deprotection followed by ester deprotection. A throughdiscussion of the precedures, reagents and conditions for removingprotecting groups is described in literature, for example, by T. W.Greene and P. G. M. Wuts in “Pretective Groups in Organic Synthesis”3^(rd) ed., John Wiley & Son, Inc., 1999.

Compound of formula (3-6) is prepared by intramolecular amide formationreaction, which is descried in the conversion of (2-5) to (2-6).

Scheme 4 illustrates a process of the invention for the preparation ofcompounds according to the invention. Reduction of the compound offormula (1-3) with a reducing agent such as, but not limited to NaBH₄,followed by protection of the amino group with Fmoc affords the compoundof formula (4-1). The reduction is carried out in a protic solvent suchas, but not limited to, methanol, ethanol, isopropanol or tert-butanolor a mixture of two protic solvents. The reaction temperature can varyfrom 0° C. to about 50° C. Protection of the amino group with Fmoc-Cl inthe presence of an organic base such as, but not limited to,triethylamine, diisopropylethylamine, DBU, N-methylmorpholine or DMAP,gives the compound of formula (4-1). The reaction is carried out in anaprotic solvent such as, but not limited to, CH₂Cl₂, DMF or THF. Thereaction temperature can vary from 0° C. to about 50° C. Furtherrearrangement of the compound of formula (4-1) in the presence of anacid, followed by acetyl protection gives the compound of formula (4-2).Suitable acids include, but are not limited to, methanesulfonic acid,toluenesulfonic acid, and camphorsulfonic acid. The rearrangementreaction is carried out in a protic solvent such as, but not limited to,methanol, ethanol, isopropanol or tert-butanol.

The acetyl protection reaction is carried out in an aprotic solvent suchas, but not limited to, CH₂Cl₂, ClCH₂CH₂Cl, DMF or THF, with aceticanhydride in the presence of base. Suitable bases include, but are notlimited to, triethylamine, diisopropylethylamine, DBU,N-methylmorpholine and DMAP. The compound of formula (4-2) is convertedto the compound of formula (4-3) with sodium methoxide in methanol.

Then the compound of formula (4-3) is coupled with a protected dipeptideof the formula (4-4) to give the compound of formula (4-5). The couplingregent can be selected from, but not limited to, DCC, EDC, di-isopropylcarbodiimide, BOP-Cl, PyBOP, PyAOP, TFFH and HATU. Suitable basesinclude, but are not limited to, triethylamine, diisopropylethylamine,DBU, N-methylmorpholine and DMAP. The coupling reaction is carried outin an aprotic solvent such as, but not limited to, CH₂Cl₂, DMF or THF.The reaction temperature can vary from 0° C. to about 50° C.

The protected dipeptides of formula (4-4) can be prepared by the methoddescribed in Hu, T. and Panek, J. S.; J. Am. Chem. Soc. 2002, 124,11372.

The compound of formula (4-5) is converted to the compound of formula(4-6) by acidic Boc deprotection. The acid can be selected from, but notlimited to, TFA, HCl in dioxane, methanesulfonic acid. A more detaileddiscussion of the procedures, reagents and conditions for removingprotecting groups is described in literature, for example, by T. W.Greene and P. G. M. Wuts in “Protective Groups in Organic Synthesis”3^(rd) ed., John Wiley & Son, Inc., 1999.

The methyl ester of compound of formula (4-6) is converted to thecorresponding acid compound of formula (4-7) via alkaline hydrolysis inprotic solvents. Representative alkali compounds include lithiumhydroxide, sodium hydroxide, potassium hydroxide, and the like. Suitablesolvents include, but are not limited to, methanol, ethanol,isopropanol, butanol, THF, 1,4-dioxane and mixtures there of. Thereaction temperature is preferably 0° to 35° C. Compound of formula(4-8) is prepared by intramolecular amide formation reaction. The regentcan be selected from, but not limited to DCC, EDC, di-isopropylcarbodiimide, BOP-Cl, PyBOP, PyAOP, TFFH and HATU. Suitable basesinclude, but are not limited to, triethylamine, diisopropylethylamine,DBU, N-methylmorpholine and DMAP. The coupling reaction is carried outin an aprotic solvent such as, but not limited to, CH₂Cl₂, DMF and THF.The reaction temperature can vary from 0° C. to about 50° C.

EXAMPLES

The compounds and processes of the present invention will be betterunderstood in connection with the following examples, which are intendedas an illustration only and not limiting of the scope of the invention.Various changes and modifications to the disclosed embodiments will beapparent to those skilled in the art and such changes and modificationsincluding, without limitation, those relating to the chemicalstructures, substituents, derivatives, formulations and/or methods ofthe invention may be made without departing from the spirit of theinvention and the scope of the appended claims.

Although the invention has been described with respect to variouspreferred embodiments, it is not intended to be limited thereto, butrather those skilled in the art will recognize that variations andmodifications may be made therein which are within the spirit of theinvention and the scope of the appended claims.

Example 1 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and R₆=Ac

Step 1a: Compound of Formula (1-1)

CsA (481 g, 0.4 mol) was dissolved in anhydrous CH₂Cl₂ (1.8 L). Aceticanhydride (163.3 g, 1.6 mol) was added followed by DMAP (48.86 g, 0.4mol) at room temperature under nitrogen. The reaction mixture wasstirred for 36 hrs. The reaction mixture was diluted with 6 L ofisopropyl acetate, followed by 8 L of water and stirred for 30 mins. Theorganic layer was separated and washed with saturated NaHCO₃ (4×6 L) andbrine (6 L). The organic phase was dried over Na₂SO₄ and concentrated.The resulted white foam was dried under vacuum to afford the compound offormula (1-1) (520 g, 95.5% HPLC purity).

MS (ESI): 1244.8 m/z (M+1).

Step 1b: Compound of Formula (1-2)

Compound of formula (1-1) (250 g, 0.2 mole) was dissolved in anhydrousCH₂Cl₂ (2 L). Trimethyloxoniumtetrafluoroborate (89.12 g, 0.6 mol) wasadded at 0° C. and the reaction mixture was stirred at room temperaturefor 20 hrs. Methanol and water (1:1 mixture, 2.5 L) was added via adropping funnel over 15 mins at 0° C. and then stirred at roomtemperature for 3 hrs. Reaction mixture was further diluted with 2 L ofCH₂Cl₂ and 2 L of water. The organic layer was separated and washed withsaturated Na₂CO₃ (2 L) and brine (2 L), and then dried over Na₂SO₄. Thesolvent was removed and the residue was purified on silica gel column toafford the compound of formula (1-2) (170 g, 92.5 HPLC purity).

MS (ESI): 1276.8 m/z (M+1).

Step 1c: Compound of Formula (1-3)

Compound of formula (1-2) (230 g, 0.18 mole) was dissolved in anhydrousTHF (1.5 L) and Phenyl thioisocyanate (24.35 g, 0.18 mole) was addedover 15 mins at 0° C. The reaction mixture was stirred at roomtemperature for 2 hrs and diluted with 1 L of water and 2.5 L of ethylacetate. The organic layer was separated and washed with brine (1 L),and then dried over Na₂SO₄ and concentrated. After dried under vacuumfor 24 hrs, the residue was dissolved in anhydrous CH₂Cl₂ (2.66 L). TFA(455 mL) was added at 0° C. over 30 mins and the reaction mixture wasstirred at room temperature for 4 hours. Reaction was quenched withsaturated Na₂CO₃ (3 L) at −15° C. The organic layer was separated andwashed with brine (3 L), and then dried over MgSO₄. Concentrated and theresidue was purified on silica gel column to afford the compound offormula (1-3) (130 g).

MS (ESI): 1149.7 m/z (M+1).

Step 1d: Compound of Formula (2-1)

Compound of formula (1-3) (11.6 g, 10 mmol)) was dissolved in anhydrousMeOH (100 ml). The solution was added sodium methoxide (1.62 g, 30 mmol)and stirred at room temperature for 6 hrs. The mixture was diluted withethyl acetate (200 ml) and quenched with 1N HCl (pH˜5). The organiclayer was separated and washed with saturated NaHCO₃ and brine, and thendried over MgSO₄. Concentrated and the residue was purified on silicagel column to afford the compound of formula (2-1) (9.8 g).

MS (ESI): 1107.8 m/z (M+1).

Step 1e: Compound of Formula (2-3)

To a 250 mL round-bottomed flask were added the compound from step 1 d(0.91 g, 0.82 mmol), the compound of formula (2-2), where R₃ is methyl,R₄ is H, R₅ is methyl and R₆ acetyl (276 mg, 1 mmol), CH₂Cl₂ (20 mL),BOP (437.9 mg, 0.99 mmol), and DMAP (241.9 mg, 1.98 mmol) respectively.The solution was stirred at room temperature for 16 hrs. Diluted withCH₂Cl₂, washed with 10% citric acid, water, saturated NaHCO₃, brine anddried over Na₂SO₄. Concentrated and the residue was purified by flashchromatography (MeOH in CH₂Cl₂, 0˜20%, v/v) to afford a white solid 1.1g.

MS (ESI): 1364.8 m/z (M+1).

Step 1f: Compound of Formula (2-4)

To a 50 mL round-bottomed flask were added the compound from step 1e(1.08 g, 0.79 mmol), THF (7 mL) and water (3 mL) respectively and thesolution was cooled to 0° C. followed by the addition of lithiumhydroxide monohydrate (100 mg, 2.4 mmol). After stirred at 0° C. for 3h, the reaction mixture was diluted withethyl acetate (50 ml), washedwith 10% citric acid solution, brine and dried over anhydrous Na₂SO₄.The solvent was removed to give the desired product 1.0 g as white foamwhich was used for next step reaction without further purification.

MS (ESI): 1350.8 m/z (M+1).

Step 1g: Compound of Formula (2-5)

To a 50 ml, round-bottomed flask were added the compound from step 1f(380 mg, 0.26 mmol) from step 1f were added CH₂Cl₂ (3 mL) and thesolution was cooled to 0° C. followed by the addition of TFA (3 mL)dropwise. The reaction mixture was stirred at 0° C. for 2 hrs and thesolvents were removed in vacuo and the residue was dissolved in CH₂Cl₂(30 mL). Washed with saturated NaHCO₃, brine and dried over anhydrousNa₂SO₄, The solvent was removed and the residue was purified by flashchromatography (MeOH/CH₂Cl₂, 1-10%, v/v) to give colorless oil 380 mg.

MS (ESI): 1250.8 m/z (M+1).

Step 1h: Compound of Formula (2-6)

To a 500 ml round-bottomed flask equipped with a dropping funnel wereadded BOP (141.5 mg, 0.32 mmol), CH₂Cl₂ (250 ml) followed by addition ofa solution of DMAP (39.1 mg, 0.32 mmol) and the compound from step 1g(200 mg, 0.16 mmol) in CH₂Cl₂ (100 mL) during 2 hrs at room temperature.The solution was stirred at room temperature for 16 hrs. The reactionwas quenched with saturated NaHCO₃. The organic layer was separated andwashed with brine dried over anhydrous Na₂SO₄. The solvent was removedand the residue was purified by flash chromatography (MeOH/CH₂Cl₂,1-10%, v/v) to give a white solid 134 mg.

MS (ESI): 1232.9 m/z (M+1).

Example 2 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and R₆=allyl

Step 2a: Compound of Formula (2-2) where R₃ is H, R₄═H, R₅═CH₃ and R₆ isAllyl

To a solution of N-Boc threonine (2.19 g, 9.98 mmol) in DMF (35 ml),sodium hydride (60% dispersion in mineral oil, 0.88 g) was added at −15°C. After stirred for 2 hrs at −15° C., allyl bromide (1.33 g, 10.98mmol) was added and the mixture was stirred for overnight at roomtemperature. The reaction mixture was poured into water (100 ml),extracted with diethyl ether (2×100 ml). The aqueous layer was acidifiedwith 10% citric acid, extracted with ethyl acetate (3×100 ml). Thecombined organic layers were washed with water (6×100 ml), then brine(2×100 ml), and dried over anhydrous Na₂SO₄. The solvent was removed andthe residue was purified by column chromatography (40% ethyl acetate inhexane) to give a pail yellow syrup 2.09 g. (yield: 80.8%).

¹H-NMR (500 MHz, CDCl₃): δ 1.23 (d, 3H, J=6.5 Hz), 1.46 (s, 9H), 3.94(dd, 1H, J=5.5, 7.0 Hz), 4.06-4.14 (m, 2H), 4.34 (dd, 1H, J=2.0, 7.0 Hz)5.17 (dd, 1H, J=0.9, 8.5 Hz), 5.23-5.27 (m, 1H), 5.30 (d, 1H, J=8.5 Hz),5.81-5.86 (m, 1H).

Step 2b: Compound of Formula (2-2) where R₃ is Methyl R₄═H, R₅═CH₃ andR₆ is Allyl

To a solution of the compound from step 2a (1.96 g, 7.57 mmol) inmixture of THF: DMF (17:1, 24 ml) was added sodium hydride (60%dispersion in mineral oil, 1.0 g) at 0° C., After stirred for 20 min at0° C. methyliodide (1.41 ml, 22.7 mmol) was added, the reaction mixturewas stirred at 0° C. for 2 hrs, then at room temperature for 17 hrs. Thereaction mixture was poured into ice water (100 ml), acidified with 10%citric acid and extracted with ethyl acetate (3×100 ml). The combinedorganic layers were washed with water (3×100 ml), brine (2×100 ml) anddried over anhydrous sodium sulfate. The solvent was removed and theresidue was purified on flash column (25% ethyl acetate inhexanes) togive pail yellow syrup (1.6 g, 77.3%).

¹H-NMR (500 MHz, CDCl₃): δ 1.20 (t, 3H, J=6.5 Hz), 1.44 (s, 3H), 1.47(s, 6H), 3.00 (d, 3H, J=7.5 Hz), 3.89-3.93 (m, 1H), 4.05-4.23 (m, 3H),4.64 (d, 0.3H J=5.0 Hz), 4.85 (d, 0.6H, J=5 Hz), 5.12 (dd, 1H, J=0.9,8.5 Hz), 5.20-5.25 (m, 1H), 5.82-5.88 (m, 1H).

Then the compound of example 2 was prepared using essentially sameprocedure from step 1e to step 1h in the preparation of the compound ofexample 1 with the compound from step 1d and the compound of formula(2-2) where R₃ is methyl and R₆ is allyl which was prepared in step 2b.

MS (ESI): 1230.7 m/z (M+1).

Example 3 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and R₆=benzyl

The compound of example 3 was prepared using essentially same procedurefrom step 1e to step 1 h in the preparation of the compound of example 1with the compound from step 1d and the compound of formula (2-2) whereR₃ is methyl and R₆ is benzyl.

MS (ESI): 1280.8 m/z (M+1).

Example 4 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 2 mL vial were added the compound of example 2 (10 mg),bromobenzene (100 μL), Pd(OAc)₂ (4.5 mg), PPh₃ (25 mg), CH₃CN (0.5 mL),Et₃N (110 μL) under N₂ and the vial was sealed and then the mixture wasirridiated on microwave reactor at 150° C. for 5 min. Solvents wereremoved and the residue was purified by column chromatography (5% MeOHin DCM) to give desired product.

ESI MS m/z=1306.91 [M+H]^(+x.)

Example 5 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Step 5a: Compound of Formula (2-2) where R₃ is Methyl R₄═H, R₅═CH₃ andR₆ is

To a 250 mL round-bottomed flask were added the compound of step 2b (1.8g, 6.59 mmol), 1,4-diacetoxy-2-butene (16.2 g, 56.2 mmol),Hoveyda-Grubbs 2nd generation catalyst (413.6 mg, 0.66 mmol) and theflask was capped with a rubber septum stopper. Vacuum was applied andbackflashed with N₂ three times. Degassed 1,2-dichloroethane (70 mL) wasadded and the reaction mixture was stirred at 40° C. for 16 hrs.Concentrated and charged with water (100 mL). The biphasic mixture wascooled to 0° C. followed by addition of 2 N NaOH (6.5 mL). The mixturewas stirred for 10 min at 0° C., extracted with Et₂O (100 mL×2). Theaqueous layer was acidified with 2 N HCl at 0° and extracted with Et₂O(250 mL×3). The combined organic layers were washed with brine, dried,concentrated to give pale brown syrup 2.2 g, 83% yield.

Then the compound of example 5 was prepared using essentially sameprocedure from step 1e to step 1h in the preparation of the compound ofexample 1 with the compound from step 1d and the compound of formula(2-2) where R₃ is methyl R₄═H, R₅═CH₃ _(—) and R₆ is

which was prepared in step 5a.

ESI MS m/z=1302.8 [M+H]⁺.

Example 6 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a solution of the compound of example 5 (22 mg), Pd₂(dba)₃ (18 mg),dppb (16 mg) in THF (2 mL) was added diethylamine (0.6 mL) under N₂. Themixture was degassed and refluxed for 4 hrs. The solvent was removed andthe residue was purified by column chromatography to give light brownfoam (20 mg).

ESIMS m/z=1315.9 [M+H]⁺.

Example 7 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 7 was prepared using essentially same procedurein the preparation of the compound of example 6 with the compound ofexample 5 and morpholine.

ESI MS m/z=1329.9 [M+H]⁺.

Example 8 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 8 was prepared using essentially same procedurein the preparation of the compound of example 6 with the compound ofexample 5 and tris-(hydroxymethyl)aminomethane.

ESI MS m/z=1363.9 [M+H]⁺.

Example 9 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a solution of compound of example 5 (47 mg, 0.038 mmol) in MeOH (2mL) was added K₂CO₃ (30 mg, 0.22 mmol) at 0° C. After stirred at 0° C.for 2 hrs, the reaction mixture was concentrated and purified by HPLC togive a white solid 25 mg.

ESI MS m/z=1260.8 [M+H]⁺.

Example 10 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a solution of compound of example 9 (330 mg, 0.26 mmol) in CH₂Cl₂ (10ml) was added Boc₂O (114 mg, 0.52 mmol), DMAP (16.5 mg) at roomtemperature. After stirred at room temperature for 4 hrs, the reactionmixture was concentrated and purified by flash chromatography to give awhite solid 300 mg.

ESI MS m/z=1360.8 [M+H]⁺.

Example 11 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 11 was prepared using essentially same procedurein the preparation of the compound of example 6 with the compound ofexample 10 and 2-morpholin-4-yl-ethanol.

ESI MS m/z=1373.9 [M+H]⁺.

Example 12 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Step 12a: Compound of Formula (2-2) where R₃ is Methyl R₄═H, R₅═CH₃ andR₆ is

To a solution of compound of formula (2-2) where R₃ is methyl, R₄ is H,R₅ is CH₃ and R₂ is

which was prepared in step 5a (2.76 g, 8 mmol) in anhydrous THF wasadded morphline (6.76 g, 80 mmol), Pd₂(dba)₂ (460 mmol, 0.5 mmol) anddppb (424 mg, 1 mmol) at room temperature. The mixture was degassed andheated to 65° C. After stirred at 65° C. for 4 hrs, the reaction mixturewas concentrated and the residue was purified by flash chromatography togive colorless oil (2.5 g).Step 12b: Compound of Formula (2-2) where R₃ is Methyl R₄═H, R₅═CH₃ andR₆ is

A mixture of compound of formula (2-2) where R₃ is methyl and R₆ is

which was prepared in step 12a (350 mg) and 10% palladium on carbon (40mg) in MeOH (10 ml) was stirred at room temperature under H₂ for 13 hrs.The mixture was filtered through a pad of celite and the filtrate wasconcentrated to give the title compound as colorless oil (350 mg).

Then the compound of example 12 was prepared using essentially sameprocedure from step 1e to step 1h in the preparation of the compound ofexample 1 with the compound from step 1d and the compound of formula(2-2) where R₃ is methyl R₄═H, R₅═CH₃ and R₆ is

which was prepared in step 12b.

ESI MS m/z=1331.9 [M+H]⁺.

Example 13 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

Step 13a: Compound of Formula (2-2) where R₃ is Methyl R₄═H, R₅═CH₃ andR₆ is

To a solution of compound of formula (2-2) where R₃ is methyl and R₄ is

which was prepared in step 5a (2.76 g, 8 mmol) in ethyl acetate (40 ml)was added 5% palladium on carbon (400 mg). The mixture was stirred underH2 (1 atm) at 0° C. for 2 hrs and filtered through a pad of celite. Thefiltrate was concentrated and purified by flash chromatography to givethe title compound as colorless oil (2.0 g).

Then the compound of example 13 was prepared using essentially sameprocedure from step 1e to step 1h in the preparation of the compound ofexample 1 with the compound from step 1d and the compound of formula(2-2) where R₃ is methyl R₄═H, R₅═CH₃ and R₆ is

which was prepared in step 13a.

ESI MS m/z=1304.8 [M+H]⁺.

Example 14 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

A solution of compound of example 13 (1.3 g, 1 mmol) in MeOH (20 mL) wasadded K₂CO₃ (152 mg, 1.1 mmol) at 0° C. After stirred at 0° C. for 2hrs, the reaction mixture was quenched with 10% citric acid (5 ml). Themixture was added saturated NaHCO₃ (50 ml) and extracted with ethylacetate (50 ml X2). The combined organic layers were washed withsaturated NaHCO₃ and brine, and then dried over anhydrous Na₂SO₄. Thesolvent was removed and the residue was purified by flash chromatographyto give the title compound as white solid (1.0 g).

ESI MS m/z=1262.8 [M+H]⁺.

Example 15 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a solution of compound of example 12 (40 mg, 0.03 mmol) in CH₂Cl₂ (1mL) was added m-CPBA (0.033 mmol) at 0° C. The reaction mixture wasstirred at 0° C. for 0.5 hr and diluted with CH₂Cl₂ (10 mL), washed with10% Na₂CO₃, brine and dried over Na₂SO₄. Filtered and the filtrate wasconcentrated, and then purified by HPLC to give the title compound aswhite solid (32 mg).

ESIMS m/z=1278.8 [M+H]⁺.

Example 16 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 4-drum vial were added compound of example 14 (100 mg, 0.079 mmol),THF (2 mL), triphenylphosphine (excess), DEAD (excess) and DPPA (excess)respectively and the solution was stirred at 50° C. overnight. The crudewas passed through a short column by eluting with a mixed solvent MeOHin CH₂Cl₂ (0˜10%) and then further purified by HPLC to afford the titlecompound (20 mg).

ESI MS m/z=1287.6 [M+H]⁺.

Example 17 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 1-drum vial were added compound of example 14 (100 mg, 0.079 mmol),CH₂Cl₂ (2 mL), CDI (25.6 mg, 0.158 mmol) respectively and the reactionmixture was stirred for 1 hr at room temperature. Morpholine (100 μL)was charged and the reaction was heated at 40° C. for 16 hrs. The crudeproduct mixture was passed through a short column by eluting with amixed solvent MeOH in CH₂Cl₂ (0˜10%) and then further purified by HPLCto afford the title compound (30 mg).

ESI MS m/z=1375.8 [M+H]⁺.

Example 18 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 18 was prepared using essential same procedurein the preparation of example 17 (30 mg).

ESI MS m/z=1319.8 [M+H]⁺.

Example 19 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

A mixture of compound of example 14 (50 mg, 0.040 mmol), Boc-Val-OH (43mg, 0.20 mmol), HATU (75 mg, 0.20 mmol), DIPEA (34.5 μL, 0.20 mmol),DMAP (4.8 mg, 0.040 mmol) in CH₂Cl₂ (1 mL) was stirred at roomtemperature for 2 hrs. The crude reaction mixture was directly purifiedon silica gel column to afford the title compound (13.6 mg).

ESI MS m/z=1461.7 [M+H]⁺.

Example 20 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 18 was prepared using essential same procedurein the preparation of example 19 (18 mg).

ESI MS m/z=1347.6 [M+H]⁺.

Example 21 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

A solution of compound of example 19 (4 mg, 0.0027 mmol) in TFA/CH₂Cl₂(2.4 mL, 1:2, v/v) was stirred at room temperature for 20 min. Thesolvent was removed and the residue was purified by flash chromatographyto afford the title compound (3.5 mg).

ESI MS m/z=1361.6 [M+H]⁺.

Example 22 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 1-drum vial were added the compound of example 14 (20 mg, 0.016mmol), THF (1 mL), triphenylphosphine (55 mg, 0.21 mmol), DEAD (32.6 μL,0.21 mmol) and acetone cyanohydrin (43.9 μL, 0.48 mmol) respectively andthe solution was stirred for 0.5 h at 50° C. The crude product mixturewas passed through a short column by eluting with a mixed solvent MeOHin CH₂Cl₂ (0˜10%). Prep TLC purification afforded the title compound aswhite foam (12 mg).

ESI MS m/z=1271.8 [M+H]⁺.

Example 23 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 1-drum vial were added the compound of example 14 (20 mg, 0.016mmol), THF (1 mL), triphenylphosphine (55 mg, 0.21 mmol), DEAD (32.6 μL,0.21 mmol) and tetrazole in acetonitrile (0.48 mmol) respectively andthe solution was stirred for 0.5 h at 50° C. The crude product mixturewas passed through a short column by eluting with a mixed solvent MeOHin CH₂Cl₂ (0˜10%). HPLC purification afforded the title compound aswhite foam (8 mg).

ESI MS m/z=1314.8 [M+H]⁺.

Example 24 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 1-drum vial were added the compound of example 14 (20 mg, 0.016mmol), THF (1 mL), triphenylphosphine (55 mg, 0.21 mmol), DEAD (32.6 μL,0.21 mmol) and tetrazole in acetonitrile (0.48 mmol) respectively andthe solution was stirred for 0.5 h at 50° C. The crude product mixturewas passed through a short column by eluting with a mixed solvent MeOHin CH₂Cl₂ (0˜10%). HPLC purification afforded the title compound aswhite foam (6 mg).

ESI MS m/z=1314.8 [M+H]⁺.

Example 25 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 1-drum vial were added compound of example 16 (50 mg, 0.039 mmol),PPh₃ (20.4 mg, 0.078 mmol), THF (1 mL) respectively and then thesolution was heated at 60° C. for 1 h. Water (14 μL, 0.78 mmol) wasadded and the reaction was heated at 60° C. for 0.5 h. Solvents wereremoved and the residue was purified by flash chromatography (MeOH/TEA,4/1) to give the title compound as white foam (46.8 mg).

ESI MS m/z=1261.4 [M+H]⁺.

Example 26 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 26 was prepared using essential same procedurein the preparation of example 17 (20 mg).

ESI MS m/z=1305.6 [M+H]⁺.

Example 27 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 1-drum vial were added triphosgene (19.2 mg, 0.065 mmol), CH₂Cl₂ (1mL), morpholine (16.6 μL, 0.19 mmol) respectively and the turbid mixturewas stirred at room temperature for 20 mins followed by addition of thecompound of example 25 (20 mg, 0.016 mmol) and DIPEA (50 μL, 0.29 mmol).The mixture was stirred at 40° C. for 1.5 hrs. Solvents were removed andthe crude product mixture was purified by HPLC to give the titlecompound as white foam (6.0 mg).

ESI MS m/z=1374.7 [M+H]⁺.

Example 28 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 1-drum vial were added compound of example 14 (100 mg, 0.079 mmol),CH₂Cl₂ (2 mL), Dess-Martin periodinane (40.3 mg, 0.095 mmol)respectively and the reaction was stirred at room temperature for 20min. Diluted with CH₂Cl₂, washed with saturated NaHCO₃, brine, and driedover Na₂SO₄. Filtered, concentrated, purified by HPLC to give the titlecompound as white foam (47 mg).

ESI MS m/z=1260.6 [M+H]⁺.

Example 29 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 1-drum vial were added the compound of example 28 (10 mg, 0.0079mmol), EtOH (1 mL), HONH₂.HCl (excess) respectively and the reaction wasstirred at room temperature for 1 hr. The crude reaction mixture wasdirectly purified by HPLC to give the title compound (5.5 mg).

ESI MS m/z=1276.6 [M+H]⁺.

Example 30 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 30 was prepared using essential same procedurein the preparation of example 29 (9 mg).

ESI MS m/z=1289.8 [M+H]⁺.

Example 31 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 31 was prepared using essential same procedurein the preparation of example 29 (10 mg).

ESI MS m/z=1351.8[M+H]⁺.

Example 32 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 32 was prepared using essential same procedurein the preparation of example 29 (10 mg).

ESI MS m/z=1365.7 [M+H]⁺.

Example 33 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 33 was prepared using essential same procedurein the preparation of example 29 (11 mg).

ESI MS m/z=1432.8 [M+H]⁺.

Example 34 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 34 was prepared using essential same procedurein the preparation of example 29 (11 mg).

ESI MS m/z=1433.8 [M+H]⁺.

Example 35 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 35 was prepared using essential same procedurein the preparation of example 29 (11 mg).

ESI MS m/z=1421.8 [M+H]⁺.

Example 36 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 36 was prepared using essential same procedurein the preparation of example 29 (11 mg).

ESI MS m/z=1464.9 [M+H]⁺.

Example 37 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 1-drum vial were added the compound of example 28 (10 mg, 0.0079mmol), EtOH (1 mL), Et₃N.HCl (excess) respectively and the reaction wasstirred at room temperature for 1 hr. The crude reaction mixture wasdirectly purified by HPLC to give the title compound (8.5 mg).

ESI MS m/z=1334.9 [M+H]⁺.

Example 38 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 38 was prepared using essential same procedurein the preparation of example 29 (11 mg).

ESI MS m/z=1464.9 [M+H]⁺.

Example 39 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

A solution of the compound of example 39 (10 mg, 0.0076 mmol) in 7 N NH₃in MeOH (2 mL) was stirred for 0.5 hr at room temperature. Concentratedand purified by silica gel column (7% MeOH in CH₂Cl₂) to give the titlecompound as white solid (6.0 mg).

ESI MS m/z=1278.8 [M+H]⁺.

Example 40 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 2-drum vial were added the compound of example 14 (100 mg, 0.079mmol), CH₂Cl₂ (1 mL), TEA (48.4 μL, 0.35 mmol) respectively and thesolution was cooled to 0° C. followed by addition of mesyl chloride(13.5 μL, 0.174 mmol). The reaction was stirred at 0° C. for 2 hrs andthen quenched with water. Extracted with CH₂Cl₂, dried, concentrated anda portion of crude product 13 mg was purified by silica gel column (10%MeOH in CH₂Cl₂) to give the title compound as white powder (10 mg).

ESI MS m/z=1340.7 [M+H]⁺.

Example 41 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 1-drum vial were added the compound of example 40 (45 mg, 0.034mmol), DMF (1.5 mL) and NaSMe (excess) respectively and the reaction wasstirred at 0° C. for 0.5 hr. Diluted with CH₂Cl₂ and then quenched withacetic acid. Extracted with CH₂Cl₂, Washed with water, dried,concentrated and purified by Prep TLC to give the title compound aswhite powder (26.2 mg).

ESI MS m/z=1292.6 [M+H]⁺.

Example 42 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 2-drum vial were added the compound of example 28 (104 mg, 0.083mmol), acetonitrile (2 mL), 2-methyl-2-butene (0.5 mL), a freshlyprepared solution of 45 mg (0.4 mmol) of 80% NaClO₂, 45 mg (0.3 mmol) ofNaH₂PO₄.H₂O, and 1.5 mL of H₂O. After stirred for 15 min at 0° C., thereaction was diluted with CH₂Cl₂, washed with brine. Dried,concentrated, purified by flash chromatography eluting with MeOH/CH₂Cl₂(1˜10%) to give the title compound as white foam (70 mg).

ESI MS m/z=1276.6 [M+H]⁺.

Example 43 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

A mixture of the compound of example 42 (20 mg, 0.016 mmol), HONH₂.HCl(11 mg, 0.16 mmol), HATU (60.8 mg, 0.16 mmol), and DIPEA (56 μL, 0.32mmol) in CH₂Cl₂ (1 mL) was stirred at room temperature for 0.5 hr.Concentrated and purified by HPLC to give the title compound as whitefoam (4.4 mg).

ESI MS m/z=1291.6 [M+H]⁺.

Example 44 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

The compound of example 44 was prepared using essentially the sameprocedure in the preparation of example 43 (5.0 mg).

ESI MS m/z=1305.7 [M+H]⁺.

Example 45 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 10 mL Schlenk tube were added the compound of example 14 (15 mg,0.011 mmol), CH₂Cl₂ (1 mL), and the solution was cooled to −78° C. underN₂ followed by addition of DAST (6.1 μL, 0.046 mmol). The reaction waswarmed to room temperature slowly and stirred overnight. Quenched withwater, extracted with CH₂Cl₂, dried, concentrated and purified by PrepTLC (4% MeOH in CH₂Cl₂) to give the title compound as white foam (7.0mg).

ESIMS m/z=1264.4 [M+H]⁺.

Example 46 Compound of Formula V: R₃═CH₃, R₄═H, R₅═CH₃ and

To a 1-drum vial were added the compound of example 29 (10 mg, 0.0078mmol), CH₂Cl₂ (1 mL), CDI (3.8 mg, 0.024 mmol), TEA (11 μL, 0.078 mmol)respectively and the solution was stirred at room temperature for 19hrs. Concentrated and purified by Prep TLC (5% MeOH in CH₂Cl₂) to givethe title compound as white foam (6.2 mg).

ES IMS m/z=1257.8 [M+H]⁺.

Example 47 Compound of Formula V: R₃=allyl, R₄═H, R₅═CH₃ and

Step 47a: Compound of Formula (2-2) where R₃ is H, R₄═H, R₅═CH₃ and R₆is

To a 500 mL round-bottomed flask were added the compound of formula(2-2) where R₃ is H and R₆ is allyl, which was prepared in step 2a (6.4g, 24.7 mmol), Cis-1,4-Diactoxy-2-butene (42.5 g, 247 mmol),Hoveyda-Grubbs 2^(nd) generation catalyst (770 mg, 1.23 mmol) and theflask was capped with a rubber septum stopper. Vacuum was applied andbackflashed with N₂ three times. Degassed DCE (250 mL) was added and thereaction was heated at 40° C. for 18 h. Concentrated and charged withwater (100 mL). The biphasic mixture was cooled to 0° C. followed byaddition of 2 N NaOH (12.4 mL). The mixture was stirred for 10 min at 0°C., and then transferred to a separational funnel. Water (150 mL) andEt₂O (300 mL) were added. The ether layer was discarded and the aqueouslayer was extracted again with Et₂O (200 mL). The aqueous layer waspoured into 1 L flask and acidified with 2 N HCl (20 mL) at 0° C.Extracted with Et₂O (250 mL×3) and the combined organic layers werewashed with brine, dried, concentrated and purified by flashchromatography to give the title compound as a yellow foam (6.5 g, 79.5%yield).

ESI MS m/z=332.25 [M+H]⁺.

Step 47b: Compound of Formula (2-2) where R₃ is H, R₄═H, R₅═CH₃ and R₂is

To a 100 mL round-bottomed flask were added the compound of formula(2-2) where R₃ is H and R₆ is

which was prepared in step 47a (3.4 g, 10.2 mmol), Pd₂(dba)₃ (748 mg,0.8 mmol), dppe (650 mg, 1.6 mmol), THF (50 mL), morpholine (8.7 g, 100mmol) under N₂ and the mixture was stirred at 65° C. for 2 hrs. Solventswere removed and the residue was purified by flash chromatography togive the title compound as yellow foam (3.4 g (contaminated withmorpholine).

ESI MS m/z=359.30 [M+H]⁺.

Step 47c: Compound of Formula (2-2) where R₃ is H, R₄═H, R₅═CH₃ and R₆is

A mixture of compound of formula (2-2) where R₃ is H and R₂ is

which was prepared in step 47b (1.7 g, 4.7 mmol) and 10% Pd/C (110 mg)in MeOH (50 mL) was stirred at room temperature under H₂ for 20 hrsfollowed by passing through a pad of celite. Concentrated to give thetitle compound as an oil (1.7 g, 100% yield).

ESI MS m/z=361.35 [M+H]⁺.

Step 47d: Compound of Formula (3-3) where R₃ is allyl, R₄═H, R₄═H,R₅═CH₃ and R₆ is

To a 25 mL round-bottomed flask were added the compound of formula (2-2)where R₃ is H and R₆ is

which was prepared in step 47c (238 mg, 0.66 mmol), THF (2 mL), ^(t)BuOK(78.5 mg, 0.70 mmol), Pd₂(dba)₃ (60.4 mg, 0.066 mmol), dppe (52.6 mg,0.13 mmol), and allylic tert-butoxy carbonate (418 mg, 2.64 mmol). Thesolution was stirred at 75° C. for 0.5 h. The reaction mixture was thencooled to room temperature, diluted with EtOAc, filtered through a padof celite. The filtrate was concentrated and purified by flashchromatography (Acetone/Hexane, 0-10%, v/v) to give the compound as anoil (148 mg, 51% yield).

ESI MS m/z=441.27 [M+H]⁺.

To a solution of the above compound (138 mg, 0.31 mmol) in CH₂Cl₂ (3 mL)was added TFA (2 mL) at 0° C. dropwise. After stirred at 0° C. for 2hrs, the solvents were removed in vacuo and the residue was dissolved inCH₂Cl₂. Washed with 10% Na₂CO₃, brine and dried over anhydrous Na₂SO₄,filtered, concentrated to give the title compound as yellow oil (111 mgin 100% yield).

ESI MS m/z=341.27 [M+H]⁺.

Step 47e: Compound of Formula (3-1) where R_(p) is TMS

To a solution of compound of formula (2-1) (3.5 g, 3.16 mmol) and DIPEA(1.22 g, 9.5 mmol) in 1,4-dioxane (140 mL) was added a solution of Boc₂O(1.37 g, 6.32 mmol) in 1,4-dioxane (30 mL) at 0° C. dropwise. Afterstirred at 0° C. for 10 mins and room temperature for 2 hrs, thereaction was quenched with MeOH. The solvents were removed in vacuo andthe residue was dissolved in EtOAc. Washed with saturated KHSO₄solution, brine and dried over anhydrous Na₂SO₄, the solvent was removedand the residue was purified by flash chromatography (MeOH/CH₂Cl₂,0-10%, v/v) to give the compound 3.3 g as white foam in 87% yield.

ESI MS m/z=1208.37 [M+H]⁺.

To a solution of the above compound (1.2 g, 1.0 mmol), 1-methylimidazole(246 mg, 3 mmol) and N,O-bis(trimethylsilyl)acetamide (2.03 g, 10 mmol)in CH₂Cl₂ (2 mL) was added TMSCl (107 mg, 1.0 mmol) at 0° C. dropwise.After stirred at 0° C. for 10 mins and room temperature for 1 hr, thereaction was quenched with MeOH. The solvents were removed in vacuo andthe residue was dissolved in EtOAc, washed with saturated KHSO₄solution, brine and dried over anhydrous Na₂SO₄, filtered, concentrated,and purified by flash chromatography (Hexanes/Acetone, 90-50%, v/v) togive the title compound as white foam (921 mg, 72% yield).

ESIMS m/z=1280.45 [M+H]⁺.

Step 47f: Compound of Formula (3-2)

To a solution of compound of formula (3-1) where R_(p) is TMS (0.66 g,0.52 mmol) in THF (10 mL) and water (5 mL) at 0° C. was added LiOHsolution (2.0 mmol, 4 mL 0.5 M solution in water). After stirred at 0°C. for 1 h, the reaction mixture was diluted with EtOAc, washed withsaturated KHSO₄ solution, brine and dried over anhydrous Na₂SO₄.Filtered, the filtrate was concentrated to give the title compound as awhite foam (603 mg).

ESI MS m/z=1194.21 [M+H]⁺.

Step 47g: Compound of Formula (3-4) where R₃ is Allyl and R₆ is

To a solution of compound of formula (3-2), which was prepared in step47f (500 mg, 0.40 mmol), compound of formula (3-3) where R₃ is allyl andR₆ is

which was prepared in step 47d (110 mg, 0.32 mmol) and DMAP (138 mg,1.12 mmol) in DMF (2 mL) was added HATU (307 mg, 0.81 mmol) at roomtemperature. After stirred at room temperature for 48 hrs and 40° C. for4 hrs, the reaction mixture was quenched with aqueous saturated NaHCO₃.The resulting mixture was extracted with CH₂Cl₂. The organic layer waswashed with saturated KHSO₄ solution, brine and dried over anhydrousNa₂SO₄, filtered, concentrated, and purified by flash chromatography(CH₂Cl₂/MeOH, 100-90%, v/v) to give the title compound 120 mg as whitefoam in 25% yield.

ESI MS m/z=1516.71 [M+H]⁺.

Step 47h: Compound of Formula (3-5) where R₃ is Allyl and R₆ is

To a solution of compound of formula (3-4) where R₃ is allyl and R₆ is

which was prepared in step 47g (60 mg, 0.04 mmol), Dimedone (11.2 mg,0.08 mmol) in THF (1 mL) was added Pd(Ph₃P)₄ (4.6 mg, 0.004 mmol) atroom temperature. The reaction mixture was stirred at room temperaturefor 20 mins and diluted with EtOAc. Filtered through a pad of celite,the filtrate was concentrated and the residue was purified by flashchromatography (CH₂Cl₂/MeOH, 5%, v/v) to give the compound 24 mg as anoil in 40% yield.

ESI MS m/z=1476.47 [M+H]⁺.

To a solution of the above compound (24 mg, 0.016 mmol) in CH₂Cl₂ (1 mL)was added TFA (1 mL) at 0° C. dropwise. After stirred at 0° C. for 2 h,the solvents were removed in vacuo and the residue was dissolved inCH₂Cl₂, then washed with saturated aqueous NaHCO₃ solution, brine anddried over anhydrous Na₂SO₄, filtered, concentrated to give the titlecompound as a pale yellow foam (14 mg, 64% yield).

ESI MS m/z=1376.47 [M+H]⁺.

Step 47i: Compound of Formula (3-6) where R₃ is Allyl and R₆ is

To a solution of TFFH (5.4 mg, 0.021 mmol), 2,4,6-sym-collidine (2.5 mg,0.021 mmol) in CH₂Cl₂ (50 mL) was added Compound of formula (3-5) whereR₃ is allyl and R₆ is

which was prepared in step 47h (14 mg, 0.010 mmol) in CH₂Cl₂ (20 mL) atroom temperature. The solution was stirred at room temperature for 16 h.The reaction mixture was then washed with aqueous KHSO₄, 10% aqueousNa₂CO₃, brine, dried over anhydrous Na₂SO₄, filtered, concentrated, andpurified by HPLC to give the title compound 2.2 mg as a white foam 16%yield.

ESI MS m/z=1357.54 [M+H]⁺.

Example 48 Compound of Formula V: R₃═CH₃, R₄═CH₃, R₅═H and R₆=Allyl

The compound of example 48 was prepared using essentially same procedurein the preparation of the compound of example 2 with the compound fromstep 1d and the compound of formula (2-2) where R₃═CH₃, R₄ ⁼CH₃, R₅═Hand R₆=allyl.

MS (ESI): 1230.7 m/z (M+1).

Example 49 Compound of Formula VI: R₃═CH₃, R₄═H, R₅═CH₃ and R₆═H

Step 49a: Compound of Formula (4-1)

Compound of formula (1-3) (61 g, 53 mmole) was dissolved in isopropanol(450 ml) and methanol (50 ml), and NaBH₄ (9.0 g, 266 mmole) was addedduring 1 hrs 1 at 0° C. The reaction mixture was stirred at roomtemperature for 2 hrs. Ethyl acetate (50 ml) was added and the mixturewas stirred at room temperature for 30 min and then quenched with 1N HClat 0° C. The pH of mixture was adjusted to pH˜9 by adding saturatedNaHCO₃ and Na₂CO₃. Extracted with ethyl acetate and washed withsaturated NaHCO₃ and brine. Dried over Na₂SO₄ and the solvent wasremoved. The residue was dried on vacuum to give the alcohol compound(59.7 g). Then the resulted compound was dissolved in DCM (500 ml).FmocCl (11.7 g, 45 mmole) and DIPEA (12.9 g, 100 mmol) were added at 0°C. The reaction mixture was stirred at 0° C. for 2 hrs. Diluted with DCM(500 ml) and washed with 10% citric acid, saturated NaHCO₃ and brine.Dried over Na₂SO₄ and the solvent was removed. The residue was purifiedon by silica gel column to give the compound of formula (4-1) (46 g).MS: (ESI) m/z (M+H) 1343.8.

Step 49b: Compound of Formula (4-2)

Compound of formula (4-1) (13.4 g, 10 mmole) was dissolved inisopropanol (100 ml). Methanesulfonic acid (100 mmole) was added at roomtemperature. The reaction mixture was stirred at 50° C. for 8 hrs. Thereaction mixture was condensed to ˜40 ml and was diluted with ethylacetate (500 ml) and quenched with saturated NaHCO₃. The pH of themixture was further adjusted to ˜9 by adding saturated Na₂CO₃. Theorganic layer was seperated and washed with brine. Dried over Na₂SO₄ andthe solvent was removed. The residue was dissolved in DCM (100 ml) andwas added acetic anhydride (2.04 g, 20 mmol) followed by TEA (4.04 g, 40mmol). The mixture was stirred at room temperature for 3 hrs andquenched with saturated NaHCO₃. The organic layer was separated andwashed with brine. Dried over Na₂SO₄ and concentrated. The residue waspurified on by silica gel column to give the compound of formula (4-2)(10 g). MS: (ESI) m/z (M+H) 1385.8.

Step 49c: Compound of Formula (4-3)

Compound of formula (4-2) (6.9 g, 5 mmole) was dissolved in methanol (50ml). NaOMe (2N in methanol, 25 ml) was added at room temperature. Thereaction mixture was stirred at room temperature for 18 hrs and quenchedwith saturated NaHCO₃. The pH of the mixture was further adjusted to ˜9by adding saturated Na₂CO₃. Organic layer was separated and washed withbrine. Dried over Na₂SO₄ and the solvent was removed. The residue waspurified on by silica gel column to give the compound of formula (4-3)(4.2 g). MS: (ESI) m/z (M+H) 1036.7.

Step 49d: Compound of Formula (4-5)

To a 250 mL round-bottomed flask were added the compound of formula(4-3) (1.04 g, 1.0 mmol), the compound of formula (4-4), where R₁₁ is

(475 mg, 1.1 mmol), CH₂Cl₂ (20 mL), BOP (531 mg, 1.2 mmol), and DMAP(241.9 mg, 1.98 mmol) respectively. The solution was stirred at roomtemperature for 16 hrs. Diluted with CH₂Cl₂, washed with 10% citricacid, water, saturated NaHCO₃, brine and dried over Na₂SO₄. Concentratedand the residue was purified by flash chromatography (MeOH in CH₂Cl₂,0˜20%, v/v) to afford a white solid 1.4 g.

MS (ESI): 1450.8 m/z (M+1)

Step 49e: Compound of Formula (4-6)

To a 50 mL round-bottomed flask were added the compound of formula (4-5)(725 mg, 0.50 mmol) from step 2d were added CH₂Cl₂ (3 mL) and thesolution was cooled to 0° C. followed by the addition of TFA (3 mL)dropwise. The reaction mixture was stirred at 0° C. for 2 hrs and thesolvents were removed in vacuo and the residue was dissolved in CH₂Cl₂(30 mL). Washed with saturated NaHCO₃, brine and dried over anhydrousNa₂SO₄, The solvent was removed and the residue was purified by flashchromatography (MeOH/CH₂Cl₂, 1-10%, v/v) to give colorless oil 730 mg.

MS (ESI): 1350.8 m/z (M+1)

Step 49f: Compound of Formula (4-7)

To a 50 ml, round-bottomed flask were added the compound from step 2e(675 mg, 0.5 mmol), THF (7 mL) and water (3 mL) respectively and thesolution was cooled to 0° C. followed by the addition of lithiumhydroxide monohydrate (100 mg, 2.4 mmol). After stirred at 0° C. for 3h, the reaction mixture was diluted withethyl acetate (50 ml), washedwith 10% citric acid solution, brine and dried over anhydrous Na₂SO₄.The solvent was removed to give the desired product 650 mg as white foamwhich was used for next step reaction without further purification.

MS (ESI): 1294.8 m/z (M+1)

Step 49g: Compound of Formula (4-8)

To a 500 ml round-bottomed flask equipped with a dropping funnel wereadded BOP (141.5 mg, 0.32 mmol), CH₂Cl₂ (250 ml) followed by addition ofa solution of DMAP (39.1 mg, 0.32 mmol) and the compound from step 2f(200 mg, 0.16 mmol) in CH₂Cl₂ (100 mL) during 2 hrs at room temperature.The solution was stirred at room temperature for 16 hrs. The reactionwas quenched with saturated NaHCO₃. The organic layer was separated andwashed with brine dried over anhydrous Na₂SO₄. The solvent was removedand the residue was purified by flash chromatography (MeOH/CH₂Cl₂,1-10%, v/v) to give a white solid 134 mg.

MS (ESI): 1276.9 m/z (M+1)

The compounds of Example 50 to Example 131 were synthesized using thegeneral methods described in Examples 1 to 49.

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

1. A compound represented by formula VI:

or a pharmaceutically acceptable salt or prodrug thereof, wherein R₃ ismethyl; R₄ is H; R₅ is methyl; and R₆ is substituted or unsubstitutedC₁-C₈ alkyl, substituted or unsubstituted C₂-C₈ alkenyl; or substitutedor unsubstituted C₂-C₈ alkenyl.
 2. A pharmaceutical compositioncomprising a therapeutically effective amount of a compound of claim 1or a pharmaceutically-acceptable salt, ester or prodrug thereof, incombination with a pharmaceutically acceptable carrier.
 3. A methodtreating viral infection in a subject in need thereof, comprisingadmistering to said subject a therapeutically-effective amount of apharmaceutical composition according to claim
 2. 4. The method accordingto claim 3, wherein said viral infection is selected from HCV, HBV, HAVand HIV infection.
 5. The method of claim 3 further comprisingcoadministering one or more additional anti-viral agents.
 6. The methodof claim 3, wherein said one or more additional anti-viral agents areselected from the group consisting of peg-interferon, ribavirin,viral-enzyme targeted compounds, viral-genome-targeted therapies,immunomodulatory agents and Toll-receptor agonists.
 7. The method ofclaim 6, wherein said one or more additional anti-viral agents areselected from peg-interferon, viral-enzyme targeted compounds, viralgenome-targeted therapy, immunomodulatory agents and toll-receptoragonist and combinations thereof.
 8. The method of claim 7, wherein saidviral genome-targeted therapy is selected from RNA interference andRNAi.
 9. The method of claim 7, wherein said immunomodulatory agent isselected from ribavarin and interferon.
 10. The compound of claim 1,wherein R₆ is selected from the group set forth below:


11. The compound of claim 10, wherein R₆ is selected from the group setforth below:


12. The compound of claim 10, wherein R₆ is


13. The compound of claim 1, wherein R₆ is allyl;